Efficacy of Preconditioned Adipose-Derived Stem Cells in Fat Grafting

Supplementation of Autologous Fat Grafts With Curcumin Preconditioned Adipose-Derived Stem Cells in the Treatment of Facial Contour Deformities

The aim of this study is to evaluate the comparison of therapeutic potential of curcumin preconditioned adipose derived stem cells (ASCs) enrichment fat grafting, naïve ASCs enrichment fat grafting and conventional fat grafting to correct facial contour deformities that cause aesthetic complications in patients.

Study Overview

• Status: Recruiting
• Conditions: Burn Scar; Craniofacial Microsomia; Romberg Disease; Post-Traumatic Scar (Disorder)
• Intervention / Treatment: Biological: Cultured ASCs enrichment fat grafting; Biological: Curcumin-preconditioned ASCs enrichment fat grafting; Other: Conventional fat grafting

Detailed Description

Human subcutaneous adipose tissue along informed consents will be collected in a lipoaspirate container or 20 cc B.D syringes under the sterilized conditions during the surgery. Fat aspirate collection will be based on certain selection criteria including pre-requisitioning non-diabetic, negative viral status patients. Adipose derived stem cells (ASCs) will be isolated from adipose tissue under sterile aseptic conditions. Firstly, the lipoaspirate container will be opened in the biosafety cabinet and drain off the blood carefully with the help of serological pipette. Then, fat will be washed thrice with 1X phosphate buffer saline (PBS) containing antibiotic and anti-mycotic solution, in order to get rid of blood vessels, hair and other type of connective tissue. After that, the tissue will be enzymatically digested with collagenase type I solution (1mg/ml prepared in Low Glucose Dulbecco's Modified Eagle Medium (LG-DMEM) and incubate for 45 minutes at 37°C on a shaker. After digestion, the enzyme will be inactivated by LG-DMEM augmented with 5% freshly isolated human serum and spin at 1200 rpm for 10 minutes. After centrifugation, fat will be discarded and the infranatant will be passed through 100 µm cell strainers to remove the debris. The filtrate will be centrifuged again for 1200rpm for 8 minutes. Pellet will be resuspended and plated in a sterile T-75 cm2 flask containing 10 ml LG-DMEM supplemented with 5% human serum. The flask will be placed in a humidified incubator at 37°C, 5% carbon dioxide (CO2). The media will be replenished after every third day until the cells got 80-90 % confluency. Cells of P2-P3 stage will be used in this study for immunocytochemistry, and intervention as well. For preconditioning, ASCs will be incubated with curcumin preconditioned medium for 24 h. Cell sterility and viability will be assessed prior to transplantation. For transplantation, 1,00,000 cells per ml of fat will be mixed, transferred to 10 cc syringes and injected at deformity site. Amount of fat will be predetermined by the clinician on the basis of facial asymmetry.

• Study Type: Interventional
• Enrollment (Anticipated): 24
• Phase: Phase 1

Contacts and Locations

• Study Contact: Name: Azra Mehmood, PhD; Phone Number: +92-333-4107008; Email: azra_mehmood@hotmail.com
• Study Contact Backup: Name: Amna Arif, M.Phil.; Phone Number: +92-333-4161068; Email: amnaarif.qureshi@gmail.com

Study Locations

• Pakistan
  • Punjab
    • Lahore, Punjab, Pakistan, 54550
      • Recruiting
      • Stem Cell Laboratory, Jinnah Burn and Reconstructive Surgery Center (JB&RSC)
      • Contact: Dr. Moazzam Nazeer Tarar, Plastic Surgeon MBBS/ FRCS; Phone Number: +92-300-8423139; Email: mntarar@gmail.com

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 16 years to 48 years (Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

Description

Inclusion criteria

• Age 18-50 years (male or female)
• Body-mass index 20-30 kg/m²
• Suitable for liposuction as assessed by the expected cosmetic results
• Provided signed informed consent

Exclusion criteria

• Systemic disorders include autoimmune diseases, hemorrhagic diseases, bone marrow aplasia, sepsis, cancer, and others
• Local disorders include acne, infections and others
• Risk factors include smoking, uncompensated diabetes, hypertension, and allergy to the drugs used for general anesthesia
• Microbiological contamination of either the cell culture or the transplanted tissue

Study Plan

How is the study designed?

Design Details

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

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Cultured ASCs enrichment fat grafting; Fat graft will be supplemented with naïve ASCs	Biological: Cultured ASCs enrichment fat grafting; Patients will undergo conventional fat harvesting. Harvested fat will be mixed with cultured ASCs and grafted into recipient site with 1,00,000 cells/ml injected fat
Experimental: Curcumin-preconditioned ASCs enrichment fat grafting; Fat graft will be augmented with curcumin preconditioned ASCs	Biological: Curcumin-preconditioned ASCs enrichment fat grafting; Patients will undergo conventional fat harvesting. Harvested fat will be mixed with cultured ASCs preconditioned with curcumin and grafted into recipient site with 1,00,000 cells/ml injected fat
Active Comparator: conventional fat grafting; Fat graft will not be supplemented with ex-vivo expanded ASCs	Other: Conventional fat grafting; Patients will undergo conventional fat harvesting and grafting of fat into recipient site

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Volumetric change in facial tissue by ultrasonography	Volumetric evaluation of treated area will be done by measuring the thickness of subcutaneous tissue (in millimeters) by using baseline B mode ultrasound device. Volumetric evaluation of treated area will be done by measuring the thickness of subcutaneous tissue (in millimeters) before transplantation, 1 week after transplantation then after 3, 6 and 9 months by using baseline B mode ultrasound device.	9 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Patient Assessment	Patients will be interviewed and pre-and post-operatively and digital images will be taken until 9 months of follow up.	9 months
Surgeon Assessment	Patients will be visually inspected for facial volume by two qualified plastic surgeons unaware of treatment after grafting to check whether curcumin pretreatment have effect on MSCs origin of ASCs.	9 months

Collaborators and Investigators

• Sponsor: University of the Punjab
• Collaborators: Higher Education Commission (Pakistan); Centre of Excellence in Molecular Biology, University of the Punjab, Lahore; Jinnah Burn and Reconstructive Surgery Centre, Lahore
• Investigators: Principal Investigator: Azra Mehmood, PhD, Center of Excellence in Molecular Biology (CEMB), University of the Punjab, Lahore; Study Director: Sheikh Riazuddin, PhD, Jinnah Burn and Reconstructive Surgery Center (JB&RSC), Lahore

Publications and helpful links

General Publications

• Matsumoto D, Sato K, Gonda K, Takaki Y, Shigeura T, Sato T, Aiba-Kojima E, Iizuka F, Inoue K, Suga H, Yoshimura K. Cell-assisted lipotransfer: supportive use of human adipose-derived cells for soft tissue augmentation with lipoinjection. Tissue Eng. 2006 Dec;12(12):3375-82. doi: 10.1089/ten.2006.12.3375.
• Bashir MM, Sohail M, Ahmad FJ, Choudhery MS. Preenrichment with Adipose Tissue-Derived Stem Cells Improves Fat Graft Retention in Patients with Contour Deformities of the Face. Stem Cells Int. 2019 Nov 20;2019:5146594. doi: 10.1155/2019/5146594. eCollection 2019.
• Ghufran H, Mehmood A, Azam M, Butt H, Ramzan A, Yousaf MA, Ejaz A, Tarar MN, Riazuddin S. Curcumin preconditioned human adipose derived stem cells co-transplanted with platelet rich plasma improve wound healing in diabetic rats. Life Sci. 2020 Sep 15;257:118091. doi: 10.1016/j.lfs.2020.118091. Epub 2020 Jul 12.
• D.A. Bourne, F.M. Egro, J. Bliley, I. James, G.L. Haas, E.M. Meyer, V. Donnenberg, A. Donnenberg, B. Branstetter, K. Marra, Abstract QS11: stem cell therapy enriched fat graft reconstruction of craniofacial deficits, Plastic and Reconstructive Surgery Global Open 6(4 Suppl) (2018).
• Cao Z, Li H, Wang ZH, Liang XQ. High-Density Fat Grafting Assisted Stromal Vascular Fraction Gel in Facial Deformities. J Craniofac Surg. 2022 Jan-Feb 01;33(1):108-111. doi: 10.1097/SCS.0000000000008038.
• Wang C, Long X, Si L, Chen B, Zhang Y, Sun T, Zhang X, Zhao RC, Wang X. A pilot study on ex vivo expanded autologous adipose-derived stem cells of improving fat retention in localized scleroderma patients. Stem Cells Transl Med. 2021 Aug;10(8):1148-1156. doi: 10.1002/sctm.20-0419. Epub 2021 Apr 19.

Study record dates

Study Major Dates

• Study Start (Actual): November 20, 2020
• Primary Completion (Anticipated): October 1, 2023
• Study Completion (Anticipated): December 1, 2023

Study Registration Dates

• First Submitted: October 26, 2022
• First Submitted That Met QC Criteria: November 3, 2022
• First Posted (Actual): November 9, 2022

Study Record Updates

• Last Update Posted (Actual): November 9, 2022
• Last Update Submitted That Met QC Criteria: November 3, 2022
• Last Verified: November 1, 2022

More Information

Terms related to this study

• Keywords: Fat grafting; Preconditioning; Facial deformities; Adipose derived stem cells
• Additional Relevant MeSH Terms: Nervous System Diseases; Congenital Abnormalities; Musculoskeletal Diseases; Stomatognathic Diseases; Mouth Diseases; Bone Diseases; Cranial Nerve Diseases; Craniofacial Abnormalities; Musculoskeletal Abnormalities; Bone Diseases, Developmental; Facial Nerve Diseases; Mandibulofacial Dysostosis; Craniofacial Dysostosis; Dysostoses; Goldenhar Syndrome; Facial Hemiatrophy; Physiological Effects of Drugs; Molecular Mechanisms of Pharmacological Action; Peripheral Nervous System Agents; Enzyme Inhibitors; Analgesics; Sensory System Agents; Anti-Inflammatory Agents, Non-Steroidal; Analgesics, Non-Narcotic; Anti-Inflammatory Agents; Antirheumatic Agents; Antineoplastic Agents; Curcumin
• Other Study ID Numbers: CEMB-SC01

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: Undecided

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No