Amputation With OsseointegrationOLIMPASNCT06310239
Microbiome Population Adaptation Study
An observational study of Amputation With Osseointegration, run by Walter Reed National Military Medical Center at 1 site in United States.
Trial summary
The investigators will accomplish our research aims by collecting cutaneous microbiome samples from 50 persons that have undergone the Osseointegration (OI) surgery at eight timepoints, prospectively. Read more about this study
- Who is it for?
- Ages 18 to 75 years
- Conditions the study is for
- Amputation With Osseointegration
- What is being tested?
- Collection of biologic samples and data
- How many people
- 50 participants
- Where
- 1 siteUnited States
- Study dates
- 1 Dec 2023to 30 Nov 2029
- How long will I take part?
- 24 Months
- Phase
- Observational
- Are there results?
- Not yet: the study is under way
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 75 years
- All sexes
- No healthy volunteers
- Adult, Older adult
Study population: Male and female military health care beneficiaries aged 18-75 years old have undergone an osseointegrated percutaneous prosthetic system.
Who may be able to join
- Any adult male or female that will undergo an OI procedure between the ages of 18-75.
- Defense Enrollment Eligibility Reporting System (DEERs) eligible
Who cannot join
- Inability to provide self-consent
Trial locations and contacts
1 site in 1 country. Contact the study team or a site near you to ask about taking part.
United States1 site, 1 recruiting
- Recruiting
Orhopaedic Surgery Service, Walter Reed National Military Medical Center,
Bethesda, Maryland, 20889
Angelica M Melendez-Munoz, 202-549-3229, angelica.m.melendez-munoz.ctr@health.mil
Toby L Perkins, 240-308-0548, toby.l.perkins.ctr@health.mil
About this study
Official title: Osseointegrated Limb Implant Microbiome Population Adaptation Study
The investigators will accomplish our research aims by collecting cutaneous microbiome samples from 50 persons that have undergone the Osseointegration (OI) surgery at eight timepoints, prospectively. The investigators will also collect control samples to correct for turnovers in species compositions that may naturally occur and to compare the residual limb microbiome to the sound contralateral limb. The investigators will sequence the bacterial community using universal bacterial primers. Using these sequences, The investigators will borrow from ecological theory and calculate the alpha and beta diversity. The alpha diversity will determine the species and abundance of each species that are present, while the beata diversity will allow us to compare how species assemblages and frequencies change between time points. Then, the investigators will take a phylogenetic modeling approach to determine if particular species assemblages correlate with rates of wound healing. The investigators will construct phylogenies from the sequences at the different time points and "paint" the rate of wound healing along the phylogeny (e.g., improved, stagnated, worsened). Using Akaike and Bayesian information criterion, the investigators can determine which phylogenetic model best explains the patterns the investigators see across patients. Lastly, the investigators will quantify soft tissue stability and health and correlate this with the homeostasis of the microbial community. Specifically, the investigators will determine if redundant soft tissue leads to altered microbial communities that can impact the rate of wound healing. Finally, the investigators will further stratify these data to compare microbial communities between the sexes, upper versus lower limbs, and proximal versus distal amputations. This work will allow us to better treat infections after OI surgery and can shed light on wound healing process so that the investigators can better treat limb loss patients and the military community as a whole.
Read the detailed description
Patients that are undergoing to have OI surgery will be asked if they would like to participate in the research study. All the data collected for the study is research related and will be collected on the case report forms ( CRFs) /source documents and will be entered into the Henry Jackson Foundation (HJF) REDCap.
Below is a breakdown of the research-related study procedures. All of these procedures will be done for research purposes, none of them are for the standard of care. The investigators will coincide study visit with clinical visits/ follow ups. The investigators will collect the following information from the patients/clinician:
Baseline/Pre-operative Surgery Stage I
Demographics (DOB, sex, height, weight, BMI, length of residual limb, amputated side, side indicated for surgery, date of amputation, reason for amputation, details of reason for amputation, issues with prior/current prosthesis) Medication: The investigators will collect detailed information about the medications (names, indication, dose, frequency, route. etc.) that the patients are currently taking (including over-the counter medications, vitamins, or herbal treatment).
Hygiene Questionnaire Biome sample (Osseointegrated and control site) Flora Data Collection Sample Questionnaire Flora Data Collection Sample Questionnaire-Control Holger Score System Questionnaire ( It is a score that the investigators is given after they evaluate the participant's skin condition).
Clinical Assessment
Pre-operative Surgery Stage II, Post-Operative 3 Weeks, 3 Months, 6 Months, and 12 Months Follow-Ups from Surgery Stage II
Medication: The patients will be asked about any changes to the medications that they are taking currently.
Hygiene Questionnaire Biome sample (Osseointegrated and control site) Flora Data Collection Sample Questionnaire Flora Data Collection Sample Questionnaire-Control Holger Score System Questionnaire
Post-Operative Surgery Stage II (in the OR)
Flora Data Collection Sample Questionnaire Flora Data Collection Sample Questionnaire-Control Biome sample (Osseointegrated and control site) Holger Score System Questionnaire
24 Months Post-Operative Surgery Stage II
Medication: The patients will be asked about any changes to the medications that they are taking currently.
Hygiene Questionnaire Biome sample (Osseointegrated and control site) Flora Data Collection Sample Questionnaire Flora Data Collection Sample Questionnaire-Control Holger Score System Questionnaire Thighplasty Questionnaire Clinical Assessment
Data collected will include flora samples collections and soft tissue envelop analysis
Flora samples collections
Biome samples will be collected /swabbed by one of the study team members. The swab area of skin or abutment will be about 2 cm x 2 cm.
Samples collecting swabbing sites For unilateral amputation patients Amputated limb: Implant surface, aperture site, the midpoint between amputated site and groin or axillary crease, and amputation site Contralateral limb as a control: as same level as amputated limb (compatible location of swabbing sites to the imputed limb) For the bilateral amputation patients Both amputated limbs: Implant surface, aperture site, the midpoint between amputated site and groin or axillary crease (as control) Anytime a participant has an infection and comes in outside of research-related timepoints The investigators will collect a Biome sample (Osseointegrated and control site), Flora Data Collection Sample Questionnaire, Flora Data Collection Sample Questionnaire-Control, and clinical assessment.
Sample collection swabbing procedures
Flora is the microorganisms that exist in all living things. The investigators will use the swabbing method to collect the participant's skin biome/flora samples on their Osseointegrated and control site. The fresh swab will be in a moistening buffer such as saline or distilled water. One of the study team members will use one hand to stretch the skin site taut. With the other hand, hold the swab so the shaft is parallel to the skin surface. Apply firm pressure and rub the swab back and forth rigorously 50 times (for 30 seconds). The swab will be put in the tube buffer and will be taken to the freezer as soon as possible (within 30 min of collecting). This is a noninvasive procedure that will cause no pain but may cause discomfortable at the collecting site. If the participants ask us to stop to collection the samples because they experience any pain, The investigators will stop to collection the flora samples.
The participant's skin flora samples will be collected at Baseline/Pre-operative Surgery Stage I, Pre-operative Surgery Stage II, Immediate post-operative Surgery Stage II (in the Operation room), Post-Operative 3 Weeks, 3 Months, 6 Months, 12 Months, and 24 months Follow-Ups from Surgery Stage II.
Swab samples processing, storing, and analysis
The swab needs to go in the tube buffer and be taken to the freezer ASAP (within 30 min of collecting). These biome samples will be stored at the -20 º the Surgical Critical Care Initiative (SC2i) freezer located in building#9. Once the investigators have accumulated enough samples, the investigators will send these samples to Zymo Research Corporation for sequencing on dry ice so the samples remain frozen. After the analysis of these samples by Zymo, the results will be sent back as text and a fasta file. The text files will be the results (alpha and beta diversity, species identified, etc.) and the fasta file will be the actual genetic sequence. Zymo will dispose of all unused samples after three months once the study has been completed
The soft tissue envelope analysis
The soft tissue envelope will be measured for redundancy and scored using the Modified Holger Score System. The investigators will document soft tissue redundancy and hypermobility, drainage and irritability, and active infection. Clinical data being collected are demographics (DOB, sex, height, weight, BMI, length of residual limb, amputated side, side indicated for surgery, date of amputation, reason for amputation, details of reason for amputation, issues with prior/current prosthesis), other relevant past and present medical conditions and medications. Questionnaires that will be collected are hygiene (how many times a day do the participants clean participants' residual limb, what product(s) do the participants use to clean their residual limb, do participant apply any product(s), like lotion, after the participants have cleaned their residual limb, if yes what product(s), flora data collection sample questionnaire (date of assessment, flora sample site, flora sample control site, what site collecting from (implant surface, aperture site, contralateral limb) and person collecting sample), Holger Score System (normal skin, reddish, red and moist, granulation tissues and skin complication leading to removal of the abutment, skin overgrowth), Clinical Assessment (soft tissue-redundancy, hypermobility, drainage irritable), wound grade (improved, stagnated and worsened) and thighplasty questionnaire (did the patient have to have a thighplasty).
- Conditions
- Amputation With Osseointegration
- Being tested
- Collection of biologic samples and data
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Collection of biologic samples and dataOther
- collection of skin flora samples via swabs
- data collection regarding subject's medications See Detailed Description in Study Description section above for more details
| Study group | Role | What they receive |
|---|---|---|
| Study subjects | Individuals with a limb amputation who are undergoing an osseointegration surgery Receives: Other: Collection of biologic samples and data |
What is being measured
The primary outcome is the main question the study is designed to answer.
Determine if OI surgery causes changes in the microbiome community using phylogenies
We will use phylogenies and phylogenetic comparative methods to quantify the changes in the cutaneous microbiome flora before and after OI surgery. This will be done by comparing phylogenies across time points to determine changes in species lineage composition.
Measured: month 36
Determine if changes in the microbiome community predicts wound healing using phylogenetic comparative methods
We will use phylogenetic comparative methods and community turnover rates, quantified via alpha and beta diversity measures, to determine if community perturbations impact wound healing.
Measured: months 36-48
Quantify soft tissue health using a modified Holger Score (0 = healthy soft issue and 6 = unhealthy soft tissue) to phylogenetically correlate this with microbiome homeostasis.
We will clinically quantify soft tissue stability using a modified Holger Score, then, we will use a phylogenetic correlation to determine if soft tissue health drives microbiome homeostasis
Measured: months 36-48
Secondary outcomes (7)
Floral profiles
We will use phylogenies to build species assemblages profiles (which species are present) for each participant. Then, we will use phylogenetic methods to compare profiles within each patient through time.
Measured: months 40
Floral colonization
We will use the microbial profiles and phylogenies to determine the colonization order of the microbiome after OI surgery by comparing which species are present at baseline and which species are present at the subsequent follow-ups.
Measured: months 42
Clinical guidelines
We will design and write clinical practice guidelines to identify a skin penetration aperture infection and disseminate to the public so others can use the same guidelines
Measured: month 48
Surgical site questionnaire
We will develop and disseminate a questionnaire for the cleaning and maintenance of the skin penetration aperture to correlate hygiene routines and probability of developing a superficial infection.
Measured: month 44
Post-OI hygiene guidelines
We will develop and disseminate a skin penetration aperture hygiene guidelines based on the findings of the microbiome profiles to reduce the probability of developing a superficial infection.
Measured: month 46
Soft tissue stability guidelines
We will develop and disseminate best practice guidelines for clinicians based on our findings to educate surgeons on soft tissue stability and when surgical intervention is needed to correct soft tissue complications.
Measured: month 46
Holger score validation
We will validate our modified Holger Score (0 = healthy soft issue and 6 = unhealthy soft tissue) in osseointegrated patients for the skin penetration aperture by determining the interobserver variation.
Measured: Time Frame: month 48
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted15 Mar 2024
- Study start1 Dec 2023
- Primary completion30 Nov 2027 (estimated)
- Study completion30 Nov 2029 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (20)Papers the sponsor has linked to this study.
- Other trials for Amputation With OsseointegrationStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Other
- Time perspective
- Prospective
- Enrolment
- 50 participants (estimated)
- Target duration
- 24 Months
Sponsor and investigators
- Lead sponsor
- Walter Reed National Military Medical Center
- Sponsor type
- US federal agency
- Responsible party
- Sponsor
- Investigators
- Ean R Saberski, MD, Principal investigator, Walter Reed National Military Medical Center
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT06310239
- Sponsor's protocol ID
- WRNMMC-2023-0434
- Other IDs
- 67407
- Acronym
- OLIMPAS
- Status last verified
- Aug 2025
- First submitted
- 21 Feb 2024
- Last updated
- 26 Aug 2025
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No: We will not share individual participant data (IPD) available to other researchers.
Publications
NIH Human Microbiome Portfolio Analysis Team. A review of 10 years of human microbiome research activities at the US National Institutes of Health, Fiscal Years 2007-2016. Microbiome. 2019 Feb 26;7(1):31. doi: 10.1186/s40168-019-0620-y.
PubMed 30808411Schmidt BM. Emerging Diabetic Foot Ulcer Microbiome Analysis Using Cutting Edge Technologies. J Diabetes Sci Technol. 2022 Mar;16(2):353-363. doi: 10.1177/1932296821990097. Epub 2021 Feb 12.
PubMed 33576276Zenga J, Atkinson S, Yen T, Massey B, Stadler M, Bruening J, Peppard W, Reuben M, Hayward M, Mesich B, Buchan B, Ledeboer N, Sanchez JL, Fraser R, Lin CW, Holtz ML, Awan M, Wong SJ, Puram SV, Salzman N. A phase 2 trial of a topical antiseptic bundle in head and neck cancer surgery: Effects on surgical site infection and the oral microbiome. EBioMedicine. 2022 Jul;81:104099. doi: 10.1016/j.ebiom.2022.104099. Epub 2022 Jun 4.
PubMed 35671624Zeeuwen PL, Boekhorst J, van den Bogaard EH, de Koning HD, van de Kerkhof PM, Saulnier DM, van Swam II, van Hijum SA, Kleerebezem M, Schalkwijk J, Timmerman HM. Microbiome dynamics of human epidermis following skin barrier disruption. Genome Biol. 2012 Nov 15;13(11):R101. doi: 10.1186/gb-2012-13-11-r101.
PubMed 23153041Xu Z, Hsia HC. The Impact of Microbial Communities on Wound Healing: A Review. Ann Plast Surg. 2018 Jul;81(1):113-123. doi: 10.1097/SAP.0000000000001450.
PubMed 29746280Dunyach-Remy C, Salipante F, Lavigne JP, Brunaud M, Demattei C, Yahiaoui-Martinez A, Bastide S, Palayer C, Sotto A, Gelis A. Pressure ulcers microbiota dynamics and wound evolution. Sci Rep. 2021 Sep 16;11(1):18506. doi: 10.1038/s41598-021-98073-x.
PubMed 34531517Wolcott RD, Hanson JD, Rees EJ, Koenig LD, Phillips CD, Wolcott RA, Cox SB, White JS. Analysis of the chronic wound microbiota of 2,963 patients by 16S rDNA pyrosequencing. Wound Repair Regen. 2016 Jan-Feb;24(1):163-74. doi: 10.1111/wrr.12370. Epub 2015 Dec 10.
PubMed 26463872Gardiner M, Vicaretti M, Sparks J, Bansal S, Bush S, Liu M, Darling A, Harry E, Burke CM. A longitudinal study of the diabetic skin and wound microbiome. PeerJ. 2017 Jul 20;5:e3543. doi: 10.7717/peerj.3543. eCollection 2017.
PubMed 28740749Loesche M, Gardner SE, Kalan L, Horwinski J, Zheng Q, Hodkinson BP, Tyldsley AS, Franciscus CL, Hillis SL, Mehta S, Margolis DJ, Grice EA. Temporal Stability in Chronic Wound Microbiota Is Associated With Poor Healing. J Invest Dermatol. 2017 Jan;137(1):237-244. doi: 10.1016/j.jid.2016.08.009. Epub 2016 Aug 24.
PubMed 27566400Gerzina C, Potter E, Haleem AM, Dabash S. The future of the amputees with osseointegration: A systematic review of literature. J Clin Orthop Trauma. 2020 Feb;11(Suppl 1):S142-S148. doi: 10.1016/j.jcot.2019.05.025. Epub 2019 May 31.
PubMed 31992935Grice EA, Segre JA. The skin microbiome. Nat Rev Microbiol. 2011 Apr;9(4):244-53. doi: 10.1038/nrmicro2537.
PubMed 21407241Orgel M, Aschoff HH, Sedlacek L, Graulich T, Krettek C, Roth S, Ranker A. Analysis of Stomal Bacterial Colonialization After Transcutaneous Osseointegrated Prosthetic Systems Surgery. JAMA Netw Open. 2022 Jul 1;5(7):e2223383. doi: 10.1001/jamanetworkopen.2022.23383.
PubMed 35867064Beck JP, Grogan M, Bennett BT, Jeyapalina S, Agarwal J, Bartow-McKenney C, Bugayev J, Kubiak E, Sinclair S, Grice E. Analysis of the Stomal Microbiota of a Percutaneous Osseointegrated Prosthesis: A Longitudinal Prospective Cohort Study. J Orthop Res. 2019 Dec;37(12):2645-2654. doi: 10.1002/jor.24421. Epub 2019 Aug 2.
PubMed 31317568Branemark R, Berlin O, Hagberg K, Bergh P, Gunterberg B, Rydevik B. A novel osseointegrated percutaneous prosthetic system for the treatment of patients with transfemoral amputation: A prospective study of 51 patients. Bone Joint J. 2014 Jan;96-B(1):106-13. doi: 10.1302/0301-620X.96B1.31905.
PubMed 24395320Holgers KM, Tjellstrom A, Bjursten LM, Erlandsson BE. Soft tissue reactions around percutaneous implants: a clinical study of soft tissue conditions around skin-penetrating titanium implants for bone-anchored hearing aids. Am J Otol. 1988 Jan;9(1):56-9.
PubMed 3364537
Study at a glance
From the registry record, last updated by the sponsor on 26 Aug 2025. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Osseointegrated Limb Implant Microbiome Population Adaptation Study
- ClinicalTrials.gov ID
- NCT06310239
- Acronym / sponsor ID
- OLIMPAS / WRNMMC-2023-0434
- Conditions
- Amputation With Osseointegration
- Study type
- Observational
- Recruitment status
- Recruiting
- Enrolment
- 50 participants (estimated)
- Lead sponsor
- Walter Reed National Military Medical CenterUS federal agency
- Study start
- 1 Dec 2023 (actual)
- Primary completion
- 30 Nov 2027 (estimated)
- Study completion
- 30 Nov 2029 (estimated)
- First posted
- 15 Mar 2024 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
The investigators will accomplish our research aims by collecting cutaneous microbiome samples from 50 persons that have undergone the Osseointegration (OI) surgery at eight timepoints, prospectively. The investigators will also collect control samples to correct for turnovers in species compositions that may naturally occur and to compare the residual limb microbiome to the sound contralateral limb. The investigators will sequence the bacterial community using universal bacterial primers. Using these sequences, The investigators will borrow from ecological theory and calculate the alpha and beta diversity. The alpha diversity will determine the species and abundance of each species that are present, while the beata diversity will allow us to compare how species assemblages and frequencies change between time points. Then, the investigators will take a phylogenetic modeling approach to determine if particular species assemblages correlate with rates of wound healing. The investigators will construct phylogenies from the sequences at the different time points and "paint" the rate of wound healing along the phylogeny (e.g., improved, stagnated, worsened). Using Akaike and Bayesian information criterion, the investigators can determine which phylogenetic model best explains the patterns the investigators see across patients. Lastly, the investigators will quantify soft tissue stability and health and correlate this with the homeostasis of the microbial community. Specifically, the investigators will determine if redundant soft tissue leads to altered microbial communities that can impact the rate of wound healing. Finally, the investigators will further stratify these data to compare microbial communities between the sexes, upper versus lower limbs, and proximal versus distal amputations. This work will allow us to better treat infections after OI surgery and can shed light on wound healing process so that the investigators can better treat limb loss patients and the military community as a whole.
Detailed description
Patients that are undergoing to have OI surgery will be asked if they would like to participate in the research study. All the data collected for the study is research related and will be collected on the case report forms ( CRFs) /source documents and will be entered into the Henry Jackson Foundation (HJF) REDCap.
Below is a breakdown of the research-related study procedures. All of these procedures will be done for research purposes, none of them are for the standard of care. The investigators will coincide study visit with clinical visits/ follow ups. The investigators will collect the following information from the patients/clinician:
Baseline/Pre-operative Surgery Stage I
Demographics (DOB, sex, height, weight, BMI, length of residual limb, amputated side, side indicated for surgery, date of amputation, reason for amputation, details of reason for amputation, issues with prior/current prosthesis) Medication: The investigators will collect detailed information about the medications (names, indication, dose, frequency, route. etc.) that the patients are currently taking (including over-the counter medications, vitamins, or herbal treatment).
Hygiene Questionnaire Biome sample (Osseointegrated and control site) Flora Data Collection Sample Questionnaire Flora Data Collection Sample Questionnaire-Control Holger Score System Questionnaire ( It is a score that the investigators is given after they evaluate the participant's skin condition).
Clinical Assessment
Pre-operative Surgery Stage II, Post-Operative 3 Weeks, 3 Months, 6 Months, and 12 Months Follow-Ups from Surgery Stage II
Medication: The patients will be asked about any changes to the medications that they are taking currently.
Hygiene Questionnaire Biome sample (Osseointegrated and control site) Flora Data Collection Sample Questionnaire Flora Data Collection Sample Questionnaire-Control Holger Score System Questionnaire
Post-Operative Surgery Stage II (in the OR)
Flora Data Collection Sample Questionnaire Flora Data Collection Sample Questionnaire-Control Biome sample (Osseointegrated and control site) Holger Score System Questionnaire
24 Months Post-Operative Surgery Stage II
Medication: The patients will be asked about any changes to the medications that they are taking currently.
Hygiene Questionnaire Biome sample (Osseointegrated and control site) Flora Data Collection Sample Questionnaire Flora Data Collection Sample Questionnaire-Control Holger Score System Questionnaire Thighplasty Questionnaire Clinical Assessment
Data collected will include flora samples collections and soft tissue envelop analysis
Flora samples collections
Biome samples will be collected /swabbed by one of the study team members. The swab area of skin or abutment will be about 2 cm x 2 cm.
Samples collecting swabbing sites For unilateral amputation patients Amputated limb: Implant surface, aperture site, the midpoint between amputated site and groin or axillary crease, and amputation site Contralateral limb as a control: as same level as amputated limb (compatible location of swabbing sites to the imputed limb) For the bilateral amputation patients Both amputated limbs: Implant surface, aperture site, the midpoint between amputated site and groin or axillary crease (as control) Anytime a participant has an infection and comes in outside of research-related timepoints The investigators will collect a Biome sample (Osseointegrated and control site), Flora Data Collection Sample Questionnaire, Flora Data Collection Sample Questionnaire-Control, and clinical assessment.
Sample collection swabbing procedures
Flora is the microorganisms that exist in all living things. The investigators will use the swabbing method to collect the participant's skin biome/flora samples on their Osseointegrated and control site. The fresh swab will be in a moistening buffer such as saline or distilled water. One of the study team members will use one hand to stretch the skin site taut. With the other hand, hold the swab so the shaft is parallel to the skin surface. Apply firm pressure and rub the swab back and forth rigorously 50 times (for 30 seconds). The swab will be put in the tube buffer and will be taken to the freezer as soon as possible (within 30 min of collecting). This is a noninvasive procedure that will cause no pain but may cause discomfortable at the collecting site. If the participants ask us to stop to collection the samples because they experience any pain, The investigators will stop to collection the flora samples.
The participant's skin flora samples will be collected at Baseline/Pre-operative Surgery Stage I, Pre-operative Surgery Stage II, Immediate post-operative Surgery Stage II (in the Operation room), Post-Operative 3 Weeks, 3 Months, 6 Months, 12 Months, and 24 months Follow-Ups from Surgery Stage II.
Swab samples processing, storing, and analysis
The swab needs to go in the tube buffer and be taken to the freezer ASAP (within 30 min of collecting). These biome samples will be stored at the -20 º the Surgical Critical Care Initiative (SC2i) freezer located in building#9. Once the investigators have accumulated enough samples, the investigators will send these samples to Zymo Research Corporation for sequencing on dry ice so the samples remain frozen. After the analysis of these samples by Zymo, the results will be sent back as text and a fasta file. The text files will be the results (alpha and beta diversity, species identified, etc.) and the fasta file will be the actual genetic sequence. Zymo will dispose of all unused samples after three months once the study has been completed
The soft tissue envelope analysis
The soft tissue envelope will be measured for redundancy and scored using the Modified Holger Score System. The investigators will document soft tissue redundancy and hypermobility, drainage and irritability, and active infection. Clinical data being collected are demographics (DOB, sex, height, weight, BMI, length of residual limb, amputated side, side indicated for surgery, date of amputation, reason for amputation, details of reason for amputation, issues with prior/current prosthesis), other relevant past and present medical conditions and medications. Questionnaires that will be collected are hygiene (how many times a day do the participants clean participants' residual limb, what product(s) do the participants use to clean their residual limb, do participant apply any product(s), like lotion, after the participants have cleaned their residual limb, if yes what product(s), flora data collection sample questionnaire (date of assessment, flora sample site, flora sample control site, what site collecting from (implant surface, aperture site, contralateral limb) and person collecting sample), Holger Score System (normal skin, reddish, red and moist, granulation tissues and skin complication leading to removal of the abutment, skin overgrowth), Clinical Assessment (soft tissue-redundancy, hypermobility, drainage irritable), wound grade (improved, stagnated and worsened) and thighplasty questionnaire (did the patient have to have a thighplasty).
Study design
- Observational model
- Other
- Time perspective
- Prospective
- Target follow-up
- 24 Months
- Number of arms
- 1
| Arm | Type | Receives |
|---|---|---|
| Study subjects | – | Other: Collection of biologic samples and dataIndividuals with a limb amputation who are undergoing an osseointegration surgery |
Who may qualify
- Ages
- 18 to 75 years
- Sex
- All
- Healthy volunteers
- Not accepted
- Age groups
- Adult, Older adult
- Study population
- Male and female military health care beneficiaries aged 18-75 years old have undergone an osseointegrated percutaneous prosthetic system.
Key inclusion criteria (2 listed)
- Any adult male or female that will undergo an OI procedure between the ages of 18-75.
- Defense Enrollment Eligibility Reporting System (DEERs) eligible
Key exclusion criteria (1 listed)
- Inability to provide self-consent
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| Collection of biologic samples and data* collection of skin flora samples via swabs * data collection regarding subject's medications See Detailed Description in Study Description section above for more details | Other | Study subjects |
Primary outcome
Determine if OI surgery causes changes in the microbiome community using phylogenies
Time frame: month 36
We will use phylogenies and phylogenetic comparative methods to quantify the changes in the cutaneous microbiome flora before and after OI surgery. This will be done by comparing phylogenies across time points to determine changes in species lineage composition.
Determine if changes in the microbiome community predicts wound healing using phylogenetic comparative methods
Time frame: months 36-48
We will use phylogenetic comparative methods and community turnover rates, quantified via alpha and beta diversity measures, to determine if community perturbations impact wound healing.
Quantify soft tissue health using a modified Holger Score (0 = healthy soft issue and 6 = unhealthy soft tissue) to phylogenetically correlate this with microbiome homeostasis.
Time frame: months 36-48
We will clinically quantify soft tissue stability using a modified Holger Score, then, we will use a phylogenetic correlation to determine if soft tissue health drives microbiome homeostasis
7 secondary outcomes
Floral profiles
Time frame: months 40
Floral colonization
Time frame: months 42
Clinical guidelines
Time frame: month 48
Surgical site questionnaire
Time frame: month 44
Post-OI hygiene guidelines
Time frame: month 46
Soft tissue stability guidelines
Time frame: month 46
Holger score validation
Time frame: Time Frame: month 48
Results and safety
This study is still under way. Results are not available in the source record yet.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 26 Aug 2025.
- Results publication: PubMed 30808411NIH Human Microbiome Portfolio Analysis Team. A review of 10 years of human microbiome research activities at the US National Institutes of Health, Fiscal Years 2007-2016. Microbiome. 2019 Feb 26;7(1):31. doi: 10.1186/s40168-019-0620-y.
- Results publication: PubMed 33576276Schmidt BM. Emerging Diabetic Foot Ulcer Microbiome Analysis Using Cutting Edge Technologies. J Diabetes Sci Technol. 2022 Mar;16(2):353-363. doi: 10.1177/1932296821990097. Epub 2021 Feb 12.
- Results publication: PubMed 35671624Zenga J, Atkinson S, Yen T, Massey B, Stadler M, Bruening J, Peppard W, Reuben M, Hayward M, Mesich B, Buchan B, Ledeboer N, Sanchez JL, Fraser R, Lin CW, Holtz ML, Awan M, Wong SJ, Puram SV, Salzman N. A phase 2 trial of a topical antiseptic bundle in head and neck cancer surgery: Effects on surgical site infection and the oral microbiome. EBioMedicine. 2022 Jul;81:104099. doi: 10.1016/j.ebiom.2022.104099. Epub 2022 Jun 4.
- Results publication: PubMed 23153041Zeeuwen PL, Boekhorst J, van den Bogaard EH, de Koning HD, van de Kerkhof PM, Saulnier DM, van Swam II, van Hijum SA, Kleerebezem M, Schalkwijk J, Timmerman HM. Microbiome dynamics of human epidermis following skin barrier disruption. Genome Biol. 2012 Nov 15;13(11):R101. doi: 10.1186/gb-2012-13-11-r101.
- Results publication: PubMed 29746280Xu Z, Hsia HC. The Impact of Microbial Communities on Wound Healing: A Review. Ann Plast Surg. 2018 Jul;81(1):113-123. doi: 10.1097/SAP.0000000000001450.
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- Results publication: PubMed 27566400Loesche M, Gardner SE, Kalan L, Horwinski J, Zheng Q, Hodkinson BP, Tyldsley AS, Franciscus CL, Hillis SL, Mehta S, Margolis DJ, Grice EA. Temporal Stability in Chronic Wound Microbiota Is Associated With Poor Healing. J Invest Dermatol. 2017 Jan;137(1):237-244. doi: 10.1016/j.jid.2016.08.009. Epub 2016 Aug 24.
- Results publication: PubMed 31992935Gerzina C, Potter E, Haleem AM, Dabash S. The future of the amputees with osseointegration: A systematic review of literature. J Clin Orthop Trauma. 2020 Feb;11(Suppl 1):S142-S148. doi: 10.1016/j.jcot.2019.05.025. Epub 2019 May 31.
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- Results publication: PubMed 35867064Orgel M, Aschoff HH, Sedlacek L, Graulich T, Krettek C, Roth S, Ranker A. Analysis of Stomal Bacterial Colonialization After Transcutaneous Osseointegrated Prosthetic Systems Surgery. JAMA Netw Open. 2022 Jul 1;5(7):e2223383. doi: 10.1001/jamanetworkopen.2022.23383.
- Individual participant dataSharing planned: No. We will not share individual participant data (IPD) available to other researchers.
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part.
Ean R Saberski, MD principal investigator
Walter Reed National Military Medical Center
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Amputation With Osseointegration, run by Walter Reed National Military Medical Center at 1 site in United States.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 2 inclusion and 1 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: looking for participants now, as of the sponsor's last update on 26 Aug 2025. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in United States. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
This study is still under way. Results are not available in the source record yet.
How can a doctor refer a patient or contact the research team?
Contact Ean R Saberski, MD (301-319-4226, Ean.saberski.mil@health.mil), or a site near the patient. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Determine if OI surgery causes changes in the microbiome community using phylogenies (time frame: month 36). The study lists 3 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT06310239, which also lists 20 linked publications.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 26 Aug 2025.