The effect of platelet-rich plasma gels on adipose mesenchymal stem cells overexpressing glia-derived neurotrophic factor

Cai Weixia; Zheng Zhao; Liu Jiaqi; Liu Yang; Zhang Ting; Ji Peng; Tian Chenyang

doi:10.3760/cma.j.cn501225-20240408-00126

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > > Accepted Manuscript

Cai Weixia,Zheng Zhao,Liu Jiaqi,et al.The effect of platelet-rich plasma gels on adipose mesenchymal stem cells overexpressing glia-derived neurotrophic factor[J].Chin J Burns Wounds,2024,40(12):1-8.DOI: 10.3760/cma.j.cn501225-20240408-00126.

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The effect of platelet-rich plasma gels on adipose mesenchymal stem cells overexpressing glia-derived neurotrophic factor

doi: 10.3760/cma.j.cn501225-20240408-00126

Department of Burns and Cutaneous Surgery, Burn Center of PLA, the First Affiliated Hospital of Air Force Medical University, Xi′an710032, China

Funds:

Natural Science Foundation of Shaanxi Province of China 2022SF-399

General Program of National Natural Science Foundation of China 81471879, 82172208

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Corresponding author: Zhen Zhao, Email: zz73553@163.com
Received Date: 2024-04-08
Available Online: 2024-12-02

Abstract

Abstract

Objective To investigate the effect of platelet-rich plasma (PRP) gel on adipose mesenchymal stem cells (ADSCs) overexpressing glial cell-derived neurotrophic factor (GDNF), namely GDNF-ADSCs. Methods This study was an experimental study. Five adult male Sprague-Dawley rats (the same below) were used, and the primary ADSCs were obtained by collagenase digestion, and then the cells were indentified. The 3^rd generation ADSCs were obtained and divided into negative control group infected with empty adenovirus and overexpressing GDNF group infected with overexpressing GDNF adenovirus, according to random number table method. After 48 hours of culture, the infection of cells was observed. Five adult male Sprague-Dawley rats (the same below) were used, and the PRP was obtained after collecting blood by differential centrifugation. The microstructure of PRP was observed by scanning electron microscope. The 3^rd generation ADSCs were added into the mixture before PRP being gel and cultured for 48 h ours. The cell growth was observed by hematoxylin-eosin staining, and the cell viability/death was detected by calcein/propyl iodide staining. PRP gels loaded with overexpression of GDNF-ADSCs and PRP gels loaded with ADSCs infected with empty adenovirus were prepared. A 24-well plate/confocal dish was divided into negative control gel group and GDNF overexpressed gel group. After 48 hours of culture, the cell viability/death was detected by calcein/propyl iodide staining. After culture for 24, 48, 72 hours and 1, 2, 3, 4 weeks, the supernatant of cell culture medium was collected, the absorbance value was determined by enzyme-labeled analyzer and the GDNF content was calculated, with the sample number of 3. After 48 hours of culture, the expression of S100 protein (a specific marker of Schwann cells) was detected by immunofluorescence assay. Results After 48 hours of culture, the proportions of cells infected with adenovirus in negative control group and overexpressed GDNF group were close to 90%, and the cell growth was good. The cells in negative control group grew normally. The morphology of the cells in overexpressed GDNF group was significantly changed, about 80%-90% of the cells had two or more protruding processes, and the protruding processes were interwoven to form a network where the cells gathered. PRP gel formed a three-dimensional network structure with different pore sizes. After 48 hours of culture, ADSCs could be well attached to PRP gel, and 98% of the cells were alive. After 48 hours of culture, the cells in negative control gel group grew well and showed typical ADSC-like spindle-shaped growth. The cells in overexpressed GDNF gel group grew well, and most of the cells had two or more protrusions, and the protrusions were interwoven into a network. After culture for 24, 48, 72 hours and 1, 2, 3, 4 weeks, the content of GDNF in the supernatant of cell culture medium in overexpressed GDNF gel group was (90±10), (133±15), (150±10), (137±15), (132±18), (120±10), and (127±16) pg/mL, which was significantly higher than (41.67±7.29), (44.00±6.95), (43.00±6.09), (47.00±5.51), (49.00±5.05), (48.67±5.93), and (51.33±4.16) pg/mL in negative control gel group (with t values of 6.20, 8.08, 15.18, 9.12, 7.99, 9.61, and 7.86, respectively, P<0.05). After 48 hours of culture, the fluorescence intensity of S100 protein expression of cells in GDNF overexpressed gel group was significantly stronger than that in negative control gel group. Conclusions The prepared three-dimensional PRP gel has good biocompatibility and can lood GDNF-ADSCs and release GDNF slowly, inducing ADSCs to differentiate into Schwann cells with high S100 protein expression.
- Peripheral nerve,
- Mesenchymal stem cells,
- Glial cell derived neurotrophic factor,
- Platelet-rich plasma,
- Schwann cells,
- Adipose mesenchymal stem cells,
- Nerve repair

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