Lin Keng-Yi, Chen Poyu, Chen Alvin Chao-Yu, Chan Yi-Sheng, Lei Kin Fong, Chiu Chih-Hao
Department of Medicine, Chang Gung University, Taoyuan.
Department of Occupational Therapy and Graduate Institute of Behavioral Sciences, College of Medicine, Chang Gung University, Taoyuan.
Orthop J Sports Med. 2022 Mar 15;10(3):23259671221084706. doi: 10.1177/23259671221084706. eCollection 2022 Mar.
Rotator cuff (RC) tendinopathy is one of the most common causes of shoulder pain. Platelet-rich plasma (PRP) has been frequently used in clinical scenarios, but its efficacy remains inconsistent.
To investigate the different responses of human tenocytes from torn RCs to leukocyte-rich PRP (LR-PRP) and leukocyte-poor PRP (LP-PRP) in a 2-chamber coculture device.
Controlled laboratory study.
PRP was prepared using different platelet and leukocyte concentrations according to 5 groups: (1) LR-PRP with 5000 platelets/µL, (2) LR-PRP with 10,000 platelets/µL, (3) LP-PRP with 5000 platelets/µL, (4) LP-PRP with 10,000 platelets/µL, and (5) control with only culture medium supplementation and without PRP stimulation. Platelet-derived growth factor-AB (PDGF-AB) and transforming growth factor-β1 (TGF-β1) were measured in LR-PRP and LP-PRP via enzyme-linked immunosorbent assay. Microscopy, water-soluble tetrazolium salt assay, and quantitative real-time polymerase chain reaction were used to investigate the morphology, proliferation, and gene expression of RC tenocytes exposed to different PRP formulations. Data were collected from at least 3 independent measurements. The results were analyzed via 1-way analysis of variance, followed by the post hoc Bonferroni test.
The ratio of leukocytes to 5000 platelets/µL was 29.5 times higher in LR-PRP than in LP-PRP ( < .05). In the 5000 platelets/µL groups, the levels of TGF-β1 and PDGF-AB were both significantly higher in LR-PRP versus LP-PRP (TGF-β1: 367.0 ± 16.5 vs 308.6 ± 30.3 pg/mL, respectively [ = .043]; PDGF-AB: 172.1 ± 1.8 vs 94.1 ± 4.2 pg/mL, respectively [ < .001]). Compared with the control group, RC tenocyte proliferation was 1.42 ± 0.01 and 1.41 ± 0.03 times higher in the LR-PRP groups with 5000 platelets/µL and 10,000 platelets/µL, respectively ( < .05). The expression of tenocyte-related genes was higher in tenocytes cultured in LR-PRP.
Both the LR-PRP groups with 5000 platelets/µL and 10,000 platelets/µL induced more growth factor release and increased RC tenocyte proliferation than did the LP-PRP groups.
In RC repair, LR-PRP may be better than LP-PRP for increasing the proliferation of tenocytes.
肩袖肌腱病是肩部疼痛最常见的原因之一。富血小板血浆(PRP)已在临床中频繁使用,但其疗效仍不一致。
在双室共培养装置中研究来自撕裂肩袖的人肌腱细胞对富白细胞PRP(LR-PRP)和贫白细胞PRP(LP-PRP)的不同反应。
对照实验室研究。
根据5组不同的血小板和白细胞浓度制备PRP:(1)每微升含5000个血小板的LR-PRP,(2)每微升含10000个血小板的LR-PRP,(3)每微升含5000个血小板的LP-PRP,(4)每微升含10000个血小板的LP-PRP,以及(5)仅补充培养基且无PRP刺激的对照组。通过酶联免疫吸附测定法测量LR-PRP和LP-PRP中的血小板衍生生长因子AB(PDGF-AB)和转化生长因子-β1(TGF-β1)。使用显微镜检查、水溶性四氮唑盐测定法和定量实时聚合酶链反应来研究暴露于不同PRP制剂的肩袖肌腱细胞的形态、增殖和基因表达。数据收集自至少3次独立测量。结果通过单因素方差分析进行分析,随后进行事后Bonferroni检验。
LR-PRP中白细胞与每微升5000个血小板的比例比LP-PRP高29.5倍(P <.05)。在每微升5000个血小板的组中,LR-PRP中的TGF-β1和PDGF-AB水平均显著高于LP-PRP(TGF-β1:分别为367.0±16.5与308.6±30.3 pg/mL [P =.043];PDGF-AB:分别为172.1±1.8与94.1±4.2 pg/mL [P <.001])。与对照组相比,每微升含5000个血小板和每微升含10000个血小板的LR-PRP组中肩袖肌腱细胞增殖分别高1.42±0.01倍和1.41±0.03倍(P <.05)。在LR-PRP中培养的肌腱细胞中,肌腱细胞相关基因的表达更高。
每微升含5000个血小板和每微升含10000个血小板的LR-PRP组比LP-PRP组诱导更多生长因子释放并增加肩袖肌腱细胞增殖。
在肩袖修复中,LR-PRP在增加肌腱细胞增殖方面可能优于LP-PRP。