Objective To explore the effect of the platelet-rich plasma (PRP) on proliferation and osteogenic differentiation of the bone marrow mesenchymal stem cells (MSCs) in China goat in vitro. Methods MSCs from the bone marrow of China goat were cultured. The third passage of MSCs were treated with PRP in the PRP group (the experimental group), but the cells were cultured with only the fetal calf serum (FCS) in the FCS group (the control group). The morphology and proliferation of the cells were observed by an inverted phase contrast microscope. The effect of PRP on proliferation of MSCs was examined by the MTT assay at 2,4,6 and 8 days. Furthermore, MSCs were cultured withdexamethasone（DEX）or PRP; alkaline phosphatase (ALP) and the calcium stainingwere used to evaluate the effect of DEX or PRP on osteogenic differatiation of MSCs at 18 days. The results from the PRP group were compared with those from the FCS group. Results The time for the MSCs confluence in the PRP group was earlier than that in the FCS group when observed under the inverted phase contrast microscope. The MTT assay showed that at 2, 4, 6 and 8 days the mean absorbance values were 0.252±0.026, 0.747±0.042, 1.173±0.067, and 1.242±0.056 in the PRP group, but 0.137±0.019, 0.436±0.052, 0.939±0.036, and 1.105±0.070 in the FCS group. The mean absorbance value was significantly higher in the PRP group than in the FCS group at each observation time (P＜0.01). Compared with the FCS group, the positive-ALP cells and the calcium deposition were decreased in the PRP group; however, DEX could increase boththe number of the positiveALP cells and the calcium deposition. Conclusion The PRP can promote proliferation of the MSCs of China goats in vitro but inhibit osteogenic differentiation.
Objective To introduce the application of platelet-rich plasma (PRP) in non-vascularised bone grafts (NVBG) of maxillofacial surgery and its potential mechanism in recent years.Methods The latest articles were extensively retrieved, and the potential mechanism for PRP promotes the osteogenesis was discussed. Results PRP promotes osteogenesis when applied to NVBG, and the cytokine included in platelet is thought to be the ingredient for PRP’s effect. Some scholar has already applied PRP in the restoration of maxillofacial bone defect andgot good results. Conclusion PRP has the potential to promotesosteogenesis, and more studies are needed for further understanding of its mechanism.
Objective To investigate the effect of various concentration of platelet-rich plasma (PRP) on osteogenic differentiation of rabbit skeletal muscle-derived stem cells (SMSCs) cultured in vitro. Methods Blood drawn from the central ear arteries of 9 one-year-old New Zealand white rabbits weighing 2.5-3.0 kg (male and female) was used to prepare PRP (Landesberg method). Full blood count and platelet count in PRP were tested. Soleus muscle of right hindl imb in rabbit was obtained and used to culture SMSCs in vitro. The cells at passage 3 were randomly divided into different groups: the experimental groups in which the cells were treated by conditioned culture media with various concentrations of autologousPRP (6.25%, 12.50%, 25.00%, 50.00%), and the control group in which the cells were treated with the media without PRP. At different time points after intervention, osteogenetic activity of the cells was detected by ALP staining observation, ALP activity detection was conducted, al izarin red staining for calcium nodules and immunofluorescence staining for osteocalcin were performed, and core binding factor α1 (Cbfα1) of osteogenic gene expression was tested by RT-PCR. Results The full blood PRP count and the platelet count in PRP was (3.06 ± 0.46) × 105/μL and (18.08 ± 2.10) × 105/μL, respectively. ALP staining: the cells in all the experimental groups were positive for the staining with many black sediment particles in cytoplasm; the cells in the control group were negative staining. ALP activity: all the experimental groups were higher than the control group (P lt; 0.05), the experimental group at 12.50% was superior to other experimental groups at each time point (P lt; 0.05). Al izarin red staining: at 14 days after culture, orange-red calcium nodules were evident in all the experimental groups; no orange-red calcium nodules were observed in the control group with a mineral ization rate of zero; there were significant difference between the experimental groups and the control group in terms of mineral ization rate (P lt; 0.05), the experimental group at 12.50% had a higher mineral ization rate than other experimental groups (P lt; 0.05). Immunofluorescence staining for osteocalcin: at 7 days after culture, the experimental groups were positive for the staining with yellow fluorescence in cytoplasm, and the result of the control group was negative. RT-PCR detection: no obvious changes of the gene expression were noted at 4, 12, and 24 hoursafter culture in the control group; the gene expression in all the experimental groups was significant superior to that of control group, especially at 12 hours, and the expression in the experimental group at 12.50% was the highest. Conclusion PRP can obviously promote the osteogenic differentiation of SMSCs cultured in vitro in a concentration-dependent manner, and the 12.50% is proved to be the ideal concentration.
Objective To defect the level of platelet antibody-IgG (PA-IgG) in patients with congestive splenomegaly and hypersplenism and the change of PA-IgG level after splenectomy and subtotal splenectomy. Methods Twenty four cases of congestive splenomegaly and hypersplenism were investigated. Results The level of PA-IgG in 24 cases were higher than normal range (Ｐ＜0.01), while the platelet count were lower than normal range and there was a significant negative correlation between the level of PA-IgG and platelet count (r＝-0.4747, Ｐ＜0.05). After subtotal splenectomy or splenectomy, the level of PA-IgG descended, the platelet count raised and the negative correlation between the level of PA-IgG and platelet count disappeared. Conclusion The results suggest that there is a immunoregulation between PA-IgG and platelet. Perhaps spleen has some relation with the immunoregulation.
Objective Platelet-rich plasma (PRP) can enhance the chondrocyte prol iferation and repair of cartilage defects. To explore the safety and efficacy of intra-knee-articular injection of PRP to treat knee articular cartilage degeneration by comparing with injecting sodium hyaluronate (SH). Methods Thirty consecutive patients (30 knees) with knee articular cartilage degeneration were selected between January 2010 and June 2010. According to different injections, 30 patients wererandomly divided into PRP group (test group, n=15) and SH group (control group, n=15). There was no significant difference in gender, age, body mass index, and Kellgren-Lawrence grade between 2 groups (P gt; 0.05). Test group received 3.5 mL of PRP intra-knee-articular injections while control group received 2 mL of SH during the same time period. Both treatments were administered in series of 3 intra-knee-articular injections at 3-week intervals. Then, adverse reactions were recorded. International Knee Documentation Committee (IKDC) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and Lequesne index were used for evaluation of treatment results. Results The patients of 2 groups were followed up 6 months. There were significant differences in IKDC score, WOMAC score, and Lequesne index between pre- and post-injection in 2 groups (P lt; 0.05); no significant difference was found between different time points (3, 4, and 6 months) in test group (P gt; 0.05), while significant differences were found between the postoperative 6th month and the postoperative 3rd and 4th months in control group (P lt; 0.05). There was no significant difference in IKDC score, WOMAC score, and Lequesne index between 2 groups within 4 months (P gt; 0.05), but the effectiveness of test group was significantly better than that of control group at 6 months after injection (P lt; 0.05). Adverse reactions occurred in 12 patients (31 injections) of test group and in 12 patients (30 injections) of control group. No significant difference in onset time, termination time, and duration of adverse reactions were found between 2 groups (P gt; 0.05). Conclusion Intra-knee-articular injection of PRP to treat knee articular cartilage degeneration is safe, which can alleviate symptoms of pain and swell ing and improve the qual ity of l ife of patients; however, further data of large samples and long-term follow-up are needed to confirm the safety and effectiveness.
ObjectiveTo identify and observe the pathogenic genes and clinical phenotypes of a family with a special platelet phenotype, Hermansky-Pudlak syndrome type 6 (HSP6). MethodsA retrospective clinical study. In November 2019, one proband and three family members from six HSP families who visited Henan Eye Hospital were included in the study. The child's medical history and family history were inquired in detail. The proband and all family members underwent best corrected visual acuity (BCVA), fundus color photography, frequency-domain optical coherence tomography, and general physical examination. The proband underwent platelet transmission electron microscopy (PTEM) and colonoscopy. Peripheral venous blood was collected from the proband, her parents and younger brother, and genomic DNA was extracted. Whole exome sequencing (WES) was used to screen pathogenic genes and their loci. Bioinformatics analysis determines the pathogenicity of gene variation sites. Fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to verify the related variations. ResultsThe proband (Ⅱ-1) was a 7-year-old female. The BCVA in both eyes was 0.1, who exhibited mild horizontal nystagmus and iris depigmentation. Fundus examination revealed obvious depigmentation and an underdeveloped fovea centralis. At the age of 7, the patient underwent colonoscopy due to acute gastrointestinal bleeding. A polyp approximately 5 mm in size was found on the floor of the sigmoid colon, with erosion and mucosal leukoplakia on its surface. PTEM showed that the number of platelet dense granules was normal, but the nuclei were small or exhibited low compactness. The skin on both lower legs showed pigmentation. The clinical phenotypes of the proband’s parents (Ⅰ-1, Ⅰ-2) and younger brother (Ⅱ-2) showed no obvious abnormalities. WES revealed that the proband carried compound heterozygous variants in exon 1 of the HPS6 gene: c.60_64dup (p.L22fs) (M1) and c.1147_1148del (p.L383fs) (M2). The mother carried the M1 variant, while the father and younger brother carried the M2 variant. Bioinformatics analysis predicted that both variants were pathogenic. RT-qPCR results showed that, compared with the relative expression level of HPS6wt mRNA, the relative expression levels of HPS6L22fs and HPS6L383fs mRNA were significantly decreased (t = 3.549, 4.560; P＜0.05). Western blot analysis demonstrated that the HPS6L383fs protein was truncated, whereas the HPS6L22fs protein was not detected. ConclusionsThis family is a special HPS6 with a normal number of dense platelet granules. The compound heterozygous variations of M1 and M2 in the HPS6 gene are pathogenic genes in this family.
ObjectiveTo observe and explore the feasibility and effectiveness of platelet-rich fibrin (PRF) membrane packing and air filling in the treatment of refractory macular holes. MethodsA retrospective clinical study. From January 2019 to January 2020, 17 patients with refractory macular hole (17 eyes) who diagnosed in Renmin Hospital of Wuhan University were included in the study. Among them, there were 7 males (7 eyes) and 10 females (10 eyes), with the age of 55.18±7.91 years. All eyes underwent 23G minimally invasive vitrectomy combined with internal limiting membrane stripping and PRF packing, and air filling was performed at the end of the operation. The best corrected visual acuity (BCVA) and optical coherence tomography angiography were performed in all eyes before surgery and at 1 week and 1, 3 months after surgery. The BCVA examination was performed using a international standard logarithmic visual acuity chart, which was converted into logarithm of the minimum angle of resolution visual acuity during statistics. Taking 3 months after surgery was as the time point to judge the efficacy, the changes of BCVA, superficial retinal vascular density (SVD), foveal avascular zone (FAZ) area and central foveal thickness (CFT) before and after surgery were compared. Paired t-test was used to compare the indicators before and after surgery. ResultsAmong the 17 eyes, there were 6, 7, and 4 eyes with giant macular hole, high myopia macular hole, and recurrent macular hole, respectively; the hole diameter was 723.94±38.30 μm. Three months after surgery, all holes were closed. Compared with before surgery, the BCVA (t=4.458) and SVD (t=2.675) increased, and the CFT (t=6.329) and FAZ area (t=4.258) decreased at 3 months after surgery, and the differences were statistically significant (P<0.05). At the last follow-up, there was no complications such as intraocular hypertension and retinal detachment in all eyes.ConclusionMinimally invasive vitrectomy combined with internal limiting membrane stripping and PRF tamponade in the treatment of refractory macular holes can increase the closure rate, improve visual acuity and retinal blood perfusion.
Objective To introduce the effect of platelet-rich plasma (PRP) in the treatment of chronic tissue lesions. Methods The latest research papers concerning the relevant subject were reviewed and analyzed. Results In PRP, the platelets and cytokines, the interaction of them in the same proportion as they were in the body fluid, played promoting role in repairing the chronic damage of tissues. However, there was still way to go before PRP was appl ied for cl inical use in a widerrange. Conclusion PRP has quite a bright outlook in the treatment of chronic tissue lesions.
ObjectiveTo explore the relationship between platelet and prognosis of the total correction of tetralogy of Fallot (TOF).
MethodsWe included 11 dead patients with TOF at age of 0-6 years as a death group between 2011 and 2014 year. There were 7 male and 4 female patients at age of 18.6±16.4 months in the death group. We selected 11 matched survival patients according to age, gender of the patients as a survival group. There were also 7 male and 4 female patients at age of 19.4±16.7 months in the death group. The patients were divided into a high-platelet group and a low-platelet group. There were 6 male and 5 female patients in the high-platelet group. While there were 8 male and 3 female patients in the low-platelet group. Inflammatory cytokine and mortality were compared between the two groups.
ResultsPost-operative count of platelet in the death group was significantly lower than that in the live group, while post-operative interleukin-6 (IL-6) and C reactive protein (CRP) were much higher than those in the death group (P<0.05). Mortality was higher in the low-platelet group than that in the high-platelet group (P<0.05).
ConclusionPlatelets may alleviate inflammatory response and reduce mortality, which played a great role of protection.
ObjectiveTo investigate the effect of platelet rich plasma (PRP) in promoting wound healing of total hip arthroplasty (THA).
MethodsBetween January 2011 and January 2012, 80 patients scheduled for THA and accorded with the inclusion criteria were divided into 2 groups:wounds were treated with PRP in 40 patients (PRP group) and with normal saline in 40 patients (control group). There was no significant difference in gender, age, disease duration, injury causes, sides, fracture type, and preoperative Harris hip scores between 2 groups (P>0.05). Routine drainage and functional exercise were performed after operation.
ResultsThe postoperative drainage volume of PRP group[(137±26) mL] was significantly lower than that of control group[(424±39) mL] (t=38.726, P=0.000). At 4 days after operation, no inflammatory reaction was observed in 34 cases of PRP group and in 30 cases of control group, mild inflammatory reaction in 5 cases of PRP group and in 6 cases of control group, moderate inflammatory reaction in 1 case of PRP group and in 4 cases of control group; there was no significant difference between 2 groups (χ2=2.141, P=0.343). Wound healed by first intention in 40 patients of PRP group and in 39 patients of control group, showing no significant difference between 2 groups (P=1.000). The average follow-up period was 9 months (range, 6-12 months). The Harris hip scores of PRP group (90.2±2.5) and control group (89.3±3.1) at last follow-up were significantly better than those before operation (39.6±8.9 and 39.2±9.2 respectively) (t=34.618, P=0.000; t=32.638, P=0.000), but no significant difference was found between 2 groups (t=1.429, P=0.153).
ConclusionUsing PRP in THA wound can reduce postoperative drainage volume, improve the healing of operation incision. It is a safe, effective, and promising procedure in treatment of THA wound.
