Biological Skin Graft With Keratinocyte-stem Cell Co-cultre for Burn Patients

Study Overview
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Brief Summary
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The goal of this clinical trial is to test whether artificial skin graft can substitute autologous skin graft in current burn treatment. The main question it aims to answer is: • Can artificial skin graft result in better wound healing compared to the current burn treatment; autologous skin graft? You will: Undergo debridement surgery Receive artificial skin graft as an alternative to autologous skin graft Undergo biopsy procedure of burn area If there is a comparison group: Researchers will compare autologous skin graft group to see the wound healing process

Detailed Description
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Burn caused by working accidents occurs mostly in the male population of productive age (68.68%). Up to this date, the only treatment for grade 2B-3 burns patients in Indonesia is skin transplantation taken from their own body (autologous skin graft). This treatment has several drawbacks in which repeated surgery is sometimes necessary, the source of skin in patients with wide area of burn is limited, and patients that are in the elder age and have comorbidities have high mortality risk. In order to find an alternative for autologous skin graft therapy, a combination of tissue engineering and stem cell therapy is used to invent an artificial skin graft. The present study attempts to evaluate the efficacy of artificial skin graft seeded with autologous keratinocyte and stem cells towards wound healing in burn patients. The artificial graft used in this study is amnion bilayer; a graft that was decellularized to remove donor's cells and then layered to form into 3-D. To evaluate the efficacy, biopsies are taken from the burn area of patients to evaluate the histoarchitecture of patients' tissue by histological staining (H&E and Movat's pentachrome, IHC collagen I, collagen III, vWF, and alpha-SMA) and measure the relative gene expression by qPCR. For clinical data, burn area calculation using Rule of 9 method and thermography measurement using FLIRONE. Furthermore, systemic evaluation of patients are done by monitoring the procalcitonin, lactate, mean arterial pressure, gradation of wound, and sensory of burn area. The evaluation and measurement mentioned above are then compared to patients treated with the current available therapy; autologous skin graft. The artificial skin graft is hoped to result in equal, if not better, wound healing in burn patients compared to autologous skin graft. The investigators hope that artificial skin graft can be an option, other than autologous skin graft, in treating burn patients in the future.

Official Title
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Substitute Graft for Burns Using Biological Graft Seeded With Autologous Keratinocyte Co-cultured With Stem Cells

Conditions
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Burn Degree Second, Burn Degree Third

Intervention / Treatment
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* Procedure: Split-thickness skin graft
* Biological: Artificial skin graft
* Biological: Artificial skin graft co-culture


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Aged 18 - 55 Area of burn <50% Acute phase burn (<120 hrs) Have not undergo any surgery for burn treatment Exclusion Criteria: Immunocompromised Have comorbidities

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 55 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Sham Comparator: STSG (Split-Thickness Skin Graft)<br>Patients treated with the standard treatment; autologous skin graft | Procedure: Split-thickness skin graft<br>* Transplantation of autologous skin to burn area<br>|
| Experimental: Amnion Bilayer Only<br>Patients treated with artificial graft only | Biological: Artificial skin graft<br>* Decellularized amnion membrane formed into 3-D matrix<br>|
| Experimental: Amnion Bilayer seeded with co-culture<br>Patients treated with artificial graft seeded with autologous keratinocyte co-cultured with amnion epithelial stem cells | Biological: Artificial skin graft co-culture<br>* Decellularized amnion membrane formed into 3-D matrix seeded with autologous keratinocyte co-cultured with amnion epithelial stem cells<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Burn thickness | Thickness of burn evaluation using Rule of 9 | Day of surgery | 
| Burn thickness | Thickness of burn evaluation using Rule of 9 | Day 7 after surgery | 
| Burn thickness | Thickness of burn evaluation using Rule of 9 | Day 14 after surgery | 
| Burn thermography | Thermography evaluation using FLIRONE | Day of surgery | 
| Burn thermography | Thermography evaluation using FLIRONE | Day 7 after surgery | 
| Burn thermography | Thermography evaluation using FLIRONE | Day 14 after surgery | 
| Systemic clinical evaluation | Mean arterial pressure measurement | Day of surgery | 
| Systemic clinical evaluation | Mean arterial pressure measurement | Day 7 after surgery | 
| Systemic clinical evaluation | Mean arterial pressure measurement | Day 14 after surgery | 
| Systemic clinical evaluation | lactate measurement | Day of surgery | 
| Systemic clinical evaluation | lactate measurement | Day 7 after surgery | 
| Systemic clinical evaluation | lactate measurement | Day 14 after surgery | 
| Systemic clinical evaluation | procalcitonin measurement | Day of surgery | 
| Systemic clinical evaluation | procalcitonin measurement | Day 7 after surgery | 
| Systemic clinical evaluation | procalcitonin measurement | Day 14 after surgery | 
| Systemic clinical evaluation | urine excretion rate measurement | Day of surgery | 
| Systemic clinical evaluation | urine excretion rate measurement | Day 7 after surgery | 
| Systemic clinical evaluation | urine excretion rate measurement | Day 14 after surgery | 
| Histoarchitecture evaluation | Haematoxylin & Eosin staining | Day of surgery | 
| Histoarchitecture evaluation | Haematoxylin & Eosin staining | Day 14 after surgery | 
| Histoarchitecture evaluation | Movat's Pentachrome staining | Day of surgery | 
| Histoarchitecture evaluation | Movat's Pentachrome staining | Day 14 after surgery | 
| Immunohistochemistry | collagen-1 labelling | Day of surgery | 
| Immunohistochemistry | collagen-1 labelling | Day 14 after surgery | 
| Immunohistochemistry | collagen-3 labelling | Day of surgery | 
| Immunohistochemistry | collagen-3 labelling | Day 14 after surgery | 
| Immunohistochemistry | von Willebrand labelling | Day of surgery | 
| Immunohistochemistry | von Willebrand labelling | Day 14 after surgery | 
| Immunohistochemistry | alpha-Smooth Muscle Actin labelling | Day of surgery | 
| Immunohistochemistry | alpha-Smooth Muscle Actin labelling | Day 14 after surgery | 
| Wound healing relative gene expression | TGFB1 | Day of surgery | 
| Wound healing relative gene expression | TGFB1 | Day 14 after surgery | 
| Wound healing relative gene expression | TGFB3 | Day of surgery | 
| Wound healing relative gene expression | TGFB3 | Day 14 after surgery | 
| Wound healing relative gene expression | Wnt4 | Day of surgery | 
| Wound healing relative gene expression | Wnt4 | Day 14 after surgery | 
| Wound healing relative gene expression | CTNNB1 | Day of surgery | 
| Wound healing relative gene expression | CTNNB1 | Day 14 after surgery | 
| Wound healing relative gene expression | MMP2 | Day of surgery | 
| Wound healing relative gene expression | MMP2 | Day 14 after surgery | 
| Wound healing relative gene expression | MMP9 | Day of surgery | 
| Wound healing relative gene expression | MMP9 | Day 14 after surgery | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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burn, artificial skin graft, keratinocyte, amnion epithelial stem cells