Identification of Molecular Differences of Adipose-derived Mesenchymal Stem Cells Between Non- Responders and Responders in Treatment of Transsphincteric Perianal Fistulas Using Autologous Fat Graft Injection
Trial Snapshot
- Phase
- Not Applicable
- Status
- Completed
- Sponsor
- University of Aarhus
- Enrollment
- 27
- Primary Endpoint
- Investigation of differentiation potential of AT-MSCs to differentiate into adipocyte
Study Overview
Brief Summary
This study investigated the cellular and molecular characteristics of AT-MSCs obtained from autologous AT therapy in patients with high transphincteric perianal fistulas of crytoglandular origin. Adipose tissue was injected into anal fistulas. Characteristics of adipose tissue mesenchymal stemcells (AT-MSC) was investigated and compared in patients with fistula that healed after the treatment (responders) to patients who failed to heal (non-responders)
Detailed Description
Injection with allogene or autologous stem cells has been reported to be efficient treatment of perianal fistulas. An alternative to this treatment could be injection with freshly collected autologous adipose tissue. In this study 27 patients with cryptoglandular anal fistulas were treated with freshly collected autologous adipose tissue.A clinical assessment of the patient prior to inclusion was undertaken and a loose seton placed for at least 6 weeks prior to fat injection. An MRI of the pelvis was performed before inclusion. Fistulas with secondary tracts and/or cavities were excluded. The operation was performed in one procedure including liposuction and injection of adipose tissue. A sample of adipose tissue from all 27 patients was analyzed. AT-MSCs were isolated and characterized using cellular and molecular analyses. Clinical and MRI-scanning evaluation of fistula healing and evaluation of ano-rectal function was performed after 6 months. AT-MSCs phenotype was compared between responders and non-responders with respect to fistula healing. The evaluation of the AT-MSCs was performed in a blinded manner.
Study Design
- Study Type
- Interventional
- Allocation
- Na
- Intervention Model
- Single Group
- Primary Purpose
- Treatment
- Masking
- None
Masking Description
Characterisation of adipose tissue (AT-MSCs) was performed blinded to the result of the treatment responder/non-responder
Eligibility Criteria
- Ages
- 18 Years to 70 Years (Adult, Older Adult)
- Sex
- All
- Accepts Healthy Volunteers
- No
Inclusion Criteria
- •high trans-sphincteric fistulas
- •fistula confirmed and classified by an MRI.
- •seton (> 6 weeks) prior to fat injection
- •informed, written consent.
Exclusion Criteria
- •Anovaginal fistula
- •Active sepsis
- •IBD, immunodeficiency, prior pelvic irradiation and malignancy
- •Insulin dependent diabetes
- •More than 4 prior attempts of fistula closure
- •Tobacco smoking or nicotine substitution 8 weeks prior to fat injection.
- •Pregnancy
- •Psychiatric disorders
- •BMI ≥ 35 or BMI<20
- •Active tuberculosis
- •Patient less than 18 years
- •Unable to undergo MRI
Outcomes
Primary Outcomes
Investigation of differentiation potential of AT-MSCs to differentiate into adipocyte
Time Frame: At start of treatment
Differentiation potential of AT-MSCs: to differentiate into adipocyte measured by Oil-Red O staining and gene expression of adipogenic markers (PPARg and LPL normalized to housekeeping gene beta actin) presented as a Fold change to undifferentiated cells (arbitrary units)
Investigation of differentiation potential of AT-MSCs to differentiate into osteoblast
Time Frame: At start of treatment
Differentiation potential of AT-MSCs: to differentiate into osteoblast measured by Alizarin S staining and gene expression of osteogenic markers (BGALP and RUNX2 normalized to housekeeping gene beta actin) presented as a Fold change to undifferentiated cells (arbitrary units)
Investigation of cell proliferation of AT-MSCs
Time Frame: At start of treatment
Cell proliferation of AT-MSCs evaluated as number of cells/per day
Measurement of gene expression profile of AT-MSCs
Time Frame: At start of treatment
Gene expression of proinflammatory (NFKB, TNFa, IL1B, IL6) and senescence associated molecules(CDKN2A, TP53, TGFB1, VEGFA, IFNG, IL6) of AT-MSCs in relation to the outcome of fistula treatment (i.e. comparison between responders and non-responders). The data are normalized to housekeeping gene beta actin (arbitrary units)
Secondary Outcomes
- Evaluation of fistula healing after treatment(6 months after last injection of autologous adipose tissue)
- Functional gastroenterological outcome after treatment(6 months after last injection of autologous adipose tissue)
- Functional urological outcome after treatment(6 months after last injection of autologous adipose tissue)
- Healing of anal fistula after treatment(6 months after last injection of autologous adipose tissue)
- Defecation disorder evaluation after treatment(6 months after last injection of autologous adipose tissue)