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IRCT20181222042074N2 IRCT 2022-06-29 Mashhad University of Medical Sciences Evaluation of the effect of amniotic scaffold and keratinocyte cells on wound healing from butterfly wing disease Design and evaluation of biological dressing efficiency with tissue engineering structure in wound healing of Epidermolysis bullosa patients: the first stage of phase 2 2022-07-21 anticipated 30 Complete https://irct.ir/trial/64312 interventional Randomization: Not randomized, Blinding: Not blinded, Placebo: Not used, Assignment: Other, Purpose: Treatment. 1-2 Epidermolysis bullosa. Intervention 1: Intervention group 1: Amniotic scaffold with adipose stem cells. Isolated samples are used for liposuction to extract MSCs. Tissue tissue is treated with type 2 collagenase enzyme and the working sample is incubated with type 2 collagenase enzyme in a 37◦c shaker hot water bath for 1 hour. The container containing the 37◦c water bath is transferred under the hood. To neutralize the effect of type 2 collagenase enzyme by adding serum-containing medium (FBS 10%) and transferring the mixture to 50 ml Falcons (the tissue is seen to be changed in a 1: 3 ratio with 10% FBS culture medium and then digested into falcons. 50 ml is transferred). The suspended cells, tissue and culture medium and DPBS in the 3 supernatants in the falcons are then removed using a 25 ml pipette and the Falcons containing the cell suspension are placed in a 400G centrifuge for 6 minutes to precipitate the cell. Take. they take. The next step was to discard the supernatant and dissolve the resulting cell sediment in MSC medium and pour the cells into T75 flasks and finally transfer the flasks to the incubator (one T75 flask was used for every 15 ml of primary fat sample). The obtained cells, after completely covering the surface of the flask, obtain a cell plate and then implant it on a scaffold made of amniotic membrane. Intervention 2: Intervention group 2: Amniotic scaffold with keratinocyte cells. In this study of keratinocytes, skin samples from the abdomen or hypogastric area after liposuction surgery or neonatal circumcision samples will be used. The received skin sample is transferred under the hood and the efficient part of the skin will be cut to 10 cm x 5 cm. It will then be rinsed 3 times in PBS solution (containing 2X Penicillin / Streptomycin) and then the skin pieces will be transferred into 7 ml of Dispase solution (10 mg / ml) containing 1X Penicillin / Streptomycin and the samples will be stored at 4 ° C. The temperature will be 17-18 hours. The sample is then trypsinized for 15 minutes and then the culture medium is pipetted with 10% FBS and the cells are separated using a 70 micron mesh filter and a 400 rpm centrifuge for 6 minutes. The complete culture medium is then transferred to the flask on the cell plate and the culture medium is changed every two days. After the cells have completely filled the surface of the flask, a cell plate is taken and the cells are implanted in the amniotic scaffold. Intervention 3: Control group 1: Amniotic scaffold. Amniotic dressing will be purchased ready-made from manufacturers and the effectiveness of this cell-free scaffold in wound healing will be evaluated. Also, this type of dressing is now used to repair many cracks. Undecided - It is not yet known if there will be a plan to make this available Justification or reason for indecision in sharing IPD is Its release schedule is not yet known
public Shirin Toosi
Bu Ali Square, Health Science and Technology Park
Mashhad Iran (Islamic Republic of) 9188617871 0517034785 Toosis@mums.ac.ir Mashhad University of Medical Sciences scientific Ahmad Shah Farhat
Imam Reza Hospital
Mashhad Iran (Islamic Republic of) 0000000000 +98 51 3854 3031 FarhatA@mums.ac.ir Mashhad University of Medical Sciences Iran (Islamic Republic of) People with a diagnosis of butterfly wing genetic disease Agreeing with informed consent to participate in the study no limit 10 years Both Reluctance to cooperate Complications in wounds such as infections Q81 Epidermolysis bullosa Treatment - Devices Treatment - Devices Treatment - Devices Intervention group 1: Amniotic scaffold with adipose stem cells. Isolated samples are used for liposuction to extract MSCs. Tissue tissue is treated with type 2 collagenase enzyme and the working sample is incubated with type 2 collagenase enzyme in a 37◦c shaker hot water bath for 1 hour. The container containing the 37◦c water bath is transferred under the hood. To neutralize the effect of type 2 collagenase enzyme by adding serum-containing medium (FBS 10%) and transferring the mixture to 50 ml Falcons (the tissue is seen to be changed in a 1: 3 ratio with 10% FBS culture medium and then digested into falcons. 50 ml is transferred). The suspended cells, tissue and culture medium and DPBS in the 3 supernatants in the falcons are then removed using a 25 ml pipette and the Falcons containing the cell suspension are placed in a 400G centrifuge for 6 minutes to precipitate the cell. Take. they take. The next step was to discard the supernatant and dissolve the resulting cell sediment in MSC medium and pour the cells into T75 flasks and finally transfer the flasks to the incubator (one T75 flask was used for every 15 ml of primary fat sample). The obtained cells, after completely covering the surface of the flask, obtain a cell plate and then implant it on a scaffold made of amniotic membrane. Intervention group 2: Amniotic scaffold with keratinocyte cells. In this study of keratinocytes, skin samples from the abdomen or hypogastric area after liposuction surgery or neonatal circumcision samples will be used. The received skin sample is transferred under the hood and the efficient part of the skin will be cut to 10 cm x 5 cm. It will then be rinsed 3 times in PBS solution (containing 2X Penicillin / Streptomycin) and then the skin pieces will be transferred into 7 ml of Dispase solution (10 mg / ml) containing 1X Penicillin / Streptomycin and the samples will be stored at 4 ° C. The temperature will be 17-18 hours. The sample is then trypsinized for 15 minutes and then the culture medium is pipetted with 10% FBS and the cells are separated using a 70 micron mesh filter and a 400 rpm centrifuge for 6 minutes. The complete culture medium is then transferred to the flask on the cell plate and the culture medium is changed every two days. After the cells have completely filled the surface of the flask, a cell plate is taken and the cells are implanted in the amniotic scaffold. Control group 1: Amniotic scaffold. Amniotic dressing will be purchased ready-made from manufacturers and the effectiveness of this cell-free scaffold in wound healing will be evaluated. Also, this type of dressing is now used to repair many cracks. Wound healing rate. Timepoint: weekly. Method of measurement: Clinical examinations. Mashhad University of Medical Sciences Approved 2022-06-11 Ethics Committee of Mashhad University of Medical Sciences University St., Mashhad University of Medical Sciences (Qureshi) Office Building, Second Floor, Mashhad Vice Chancellor for Research Mashhad Razavi Khorasan Iran (Islamic Republic of)