1.Experimental study on contact co-culture of DiI labeled rat bone marrow mesenchymal stem cells and neonatal rat cardiomyocytes on polycaprolactone film to make myocardial patch
Zichang ZHANG ; Junsheng MU ; Fan ZHOU ; Ping BO ; Bin YOU
Chinese Journal of Cardiology 2024;52(5):525-531
		                        		
		                        			
		                        			Objective:To investigate the possible mechanism of DiI labeled bone marrow mesenchymal stem cells (BMSCs) in contact co-cultured with neonatal rat cardiomyocytes (CMs) on polycaprolactone (PCL) film to make myocardial patch.Methods:BMSCs from Sprague Dawley rats (aged 5-6 weeks) were isolated, cultured, and characterized for surface marker expression using flow cytometry. CMs from 15 neonatal rats were isolated and cultured. After cultured for 3 generations, BMSCs were labeled with DiI dye. On PCL film, DiI labeled BMSCs were co-cultured with CMs as the experimental group, and CMs were replaced with the same amount of unlabeled BMSCs in the control group. After 24 h of co-culture, the cell growth was observed under fluorescence microscope and the co-culture was observed under scanning electron microscope. Immunofluorescence staining was performed after 7 days to detect myocardial markers, including cardiac troponin T (cTnT) and α-actinin. BMSC differentiation on the PCL film was observed under a fluorescence microscope. The differentiation efficiency of BMSCs into cardiomyoid cells was analyzed by flow cytometry on days 1 and 7 of co-culture. Intercellular dye transfer was observed by staining CMs with calcein and co-culturing them with DiI-labeled BMSCs on PCL film. The cells were stained with immunofluorescence to detect the expression of connexin 43 (Cx43) and observe the relationship between gap junction and contact co-culture.Results:Flow cytometry showed strong positivity for CD90 and CD44 and negativity for CD11b/c and CD45 on BMSCs. After 24 h of co-culture, DiI labeled BMSCs glowed red on the PCL film, while unlabeled CMs did not; the number of cells on PCL film was large and cell morphology appeared normal under scanning electron microscope. On the 7th day of co-culture, some DiI labeled BMSCs expressed cTnT and α-actinin. Flow cytometry showed a higher differentiation rate of stem cells in the experimental group on day 7 compared to the control group ((20.12±0.15)% vs. (3.49±0.20)%, P<0.05). From the second day of co-culture, some BMSCs exhibited green dot fluorescence in Cx43 immunofluorescence staining; and by the third day, dye transfer test showed green fluorescence emission from some BMSCs. Conclusion:Contact co-culture of DiI labeled BMSCs and CMs on PCL film can make myocardial patch. The mechanism of contact co-culture promoting the differentiation and formation of myocardial patch may be associated with gap junctions and intercellular signal pathways mediated by gap junctions.
		                        		
		                        		
		                        		
		                        	
2.The jujube core-shaped tissue resection technique in alar reduction
Lehao WU ; Yihao XU ; Huan WANG ; Ruobing ZHENG ; Xulong ZHANG ; Le TIAN ; Junsheng GUO ; Shan ZHU ; Fei FAN ; Jianjun YOU
Chinese Journal of Plastic Surgery 2024;40(1):88-93
		                        		
		                        			
		                        			Objective:To investigate the clinical effect of jujube core-shaped tissue resection technique in the treatment of alar reduction.Methods:A retrospective analysis was performed for the clinical data of patients who underwent alar reduction from February 2019 to June 2022. A spindle-shaped incision line was designed along the outer edge of the base of the nasal lobule, with a width of 4-5 mm and a length of 8-12 mm. The incision line was 1-2 mm away from the alar groove, and the inner side of the design line was extended to the inside of the nasal vestibule. After the incision was made, the subcutaneous tissue was undermining dissected with curved sharp scissors, and the shape of the extended excision tissue was similar to that of jujube core. After reduction, the incision was closed with vertical mattress suture. The surgical effect and complications were followed up after surgery, and an independent attending plastic surgeon evaluated the outcome and scarring based on photos before and at the last postoperative follow-up using the global aesthetic improvement scale (GAIS) and the Vancouver scar scale (VSS). GAIS is graded as 1, 2, 3, 4, and 5 points, and the lower the score, the better the improvement(≤3 points as effective improvement). VSS includes four parameters: color (0-3 points), thickness (0-4 points), vascular distribution (0-3 points), and softness (0-5 points). The higher the score, the more significant the scar is.Results:A total of 20 patients were enrolled, including 6 males and 14 females, aged 20 to 33 years, with an average age of 24.9 years. Before the operation, there were different degrees of alar flare and alar hypertrophy. After surgery, 13 patients had mild swelling, ecchymosis, which resolved after 3 days. There were no major complications as infection, incision dehiscence, and delayed healing. After 3 to 16 months of postoperative follow-up, 2 patients gradually developed cartilage deformation and affected the appearance of the nostrils 6 to 9 months after surgery due to the combination of rib-based rhinoplasty, and the satisfactory results were obtained after revision surgery. The symmetry of the nostrils was significantly improved. The GAIS score of 20 patients was (1.20±0.41) points, of which, 16 patients were rated as 1 point and 4 patients were rated as 2 points, all of which met the improvement criteria, and the VSS score was (1.45±0.60) points, in which the average score for color, thickness, vasculature and compliance are 0.60, 0.05, 0.55 and 0.30 points respectively.Conclusion:The application of jujube core-shaped tissue resection technique for alar reduction can increase the amount of tissue removal with the same incision width, which can not only fully narrow the alar effectively but also correct the hypertrophy of the alar tissue.
		                        		
		                        		
		                        		
		                        	
3.The jujube core-shaped tissue resection technique in alar reduction
Lehao WU ; Yihao XU ; Huan WANG ; Ruobing ZHENG ; Xulong ZHANG ; Le TIAN ; Junsheng GUO ; Shan ZHU ; Fei FAN ; Jianjun YOU
Chinese Journal of Plastic Surgery 2024;40(1):88-93
		                        		
		                        			
		                        			Objective:To investigate the clinical effect of jujube core-shaped tissue resection technique in the treatment of alar reduction.Methods:A retrospective analysis was performed for the clinical data of patients who underwent alar reduction from February 2019 to June 2022. A spindle-shaped incision line was designed along the outer edge of the base of the nasal lobule, with a width of 4-5 mm and a length of 8-12 mm. The incision line was 1-2 mm away from the alar groove, and the inner side of the design line was extended to the inside of the nasal vestibule. After the incision was made, the subcutaneous tissue was undermining dissected with curved sharp scissors, and the shape of the extended excision tissue was similar to that of jujube core. After reduction, the incision was closed with vertical mattress suture. The surgical effect and complications were followed up after surgery, and an independent attending plastic surgeon evaluated the outcome and scarring based on photos before and at the last postoperative follow-up using the global aesthetic improvement scale (GAIS) and the Vancouver scar scale (VSS). GAIS is graded as 1, 2, 3, 4, and 5 points, and the lower the score, the better the improvement(≤3 points as effective improvement). VSS includes four parameters: color (0-3 points), thickness (0-4 points), vascular distribution (0-3 points), and softness (0-5 points). The higher the score, the more significant the scar is.Results:A total of 20 patients were enrolled, including 6 males and 14 females, aged 20 to 33 years, with an average age of 24.9 years. Before the operation, there were different degrees of alar flare and alar hypertrophy. After surgery, 13 patients had mild swelling, ecchymosis, which resolved after 3 days. There were no major complications as infection, incision dehiscence, and delayed healing. After 3 to 16 months of postoperative follow-up, 2 patients gradually developed cartilage deformation and affected the appearance of the nostrils 6 to 9 months after surgery due to the combination of rib-based rhinoplasty, and the satisfactory results were obtained after revision surgery. The symmetry of the nostrils was significantly improved. The GAIS score of 20 patients was (1.20±0.41) points, of which, 16 patients were rated as 1 point and 4 patients were rated as 2 points, all of which met the improvement criteria, and the VSS score was (1.45±0.60) points, in which the average score for color, thickness, vasculature and compliance are 0.60, 0.05, 0.55 and 0.30 points respectively.Conclusion:The application of jujube core-shaped tissue resection technique for alar reduction can increase the amount of tissue removal with the same incision width, which can not only fully narrow the alar effectively but also correct the hypertrophy of the alar tissue.
		                        		
		                        		
		                        		
		                        	
4.Effect of CDM3 on the co-culture of human induced pluripotent stem cells with matrigel-coated polycaprolactone to make cardiac patch
Yue DAI ; Fan ZHOU ; Jianwei ZHENG ; Junsheng MU ; Ping BO ; Bin YOU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2023;30(05):738-745
		                        		
		                        			
		                        			Objective    To provide experimental data and theoretical support for further studying the maturity of cardiac patches in other in vitro experiments and the safety in other in vivo animal experiments, through standard chemically defined and small molecule-based induction protocol (CDM3) for promoting the differentiation of human induced pluripotent stem cells (hiPSCs) into myocardium, and preliminarily preparing cardiac patches. Methods    After resuscitation, culture and identification of hiPSCs, they were inoculated on the matrigel-coated polycaprolactone (PCL). After 24 hours, the cell growth was observed by DAPI fluorescence under a fluorescence microscope, and the stemness of hiPSCs was identified by OCT4 fluorescence. After fixation, electron microscope scanning was performed to observe the cell morphology on the surface of the patch. On the 1st, 3rd, 5th, and 7th days of culture, the cell viability was determined by CCK-8 method, and the growth curve was drawn to observe the cell growth and proliferation. After co-cultured with matrigel-coated PCL for 24 hours, hiPSCs were divided into a control group and a CDM3 group, and continued to culture for 6 days. On the 8th day, the cell growth was observed by DAPI fluorescence under a fluorescence microscope, and hiPSCs stemness was identified by OCT4 fluorescence, and cTnT and α-actin for cardiomyocyte marker identification. Results    Immunofluorescence of hiPSCs co-cultured with matrigel-coated PCL for 24 hours showed that OCT4 emitted green fluorescence, and hiPSCs remained stemness on matrigel-coated PCL scaffolds. DAPI emitted blue fluorescence: cells grew clonally with uniform cell morphology. Scanning electron microscope showed that hiPSCs adhered and grew on matrigel-coated PCL, the cell outline was clearly visible, and the morphology was normal. The cell viability assay by CCK-8 method showed that hiPSCs proliferated and grew on PCL scaffolds coated with matrigel. After 6 days of culture in the control group and the CDM3 group, immunofluorescence showed that the hiPSCs in the control group highly expressed the stem cell stemness marker OCT4, but did not express the cardiac markers cTnT and α-actin. The CDM3 group obviously expressed the cardiac markers cTnT and α-actin, but did not express the stem cell stemness marker OCT4. Conclusion    hiPSCs can proliferate and grow on matrigel-coated PCL. Under the influence of CDM3, hiPSCs can be differentiated into cardiomyocyte-like cells, and the preliminary preparation of cardiac patch can provide a better treatment method for further clinical treatment of cardiac infarction.
		                        		
		                        		
		                        		
		                        	
5.Application of nasal septum extension graft drilling technique in rib-based rhinoplasty
Yihao XU ; Jianjun YOU ; Huan WANG ; Ruobing ZHENG ; Lehao WU ; Xulong ZHANG ; Le TIAN ; Junsheng GUO ; Fei FAN
Chinese Journal of Plastic Surgery 2023;39(10):1100-1105
		                        		
		                        			
		                        			Objective:To investigate the clinical application of a cartilage graft fixation method in the context of rib-based rhinoplasty.Methods:The clinical data of patients who underwent costal cartilage based rhinoplasty in the Center of Rhinoplasty and Nasal Reconstruction, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2020 to January 2022 were retrospectively analyzed. A 20 ml syringe needle was used to drill several micropores about 1 mm in diameter on the surface of the cartilage grafts prior to its placement. "septal extension grafts with micropores" were prepared, and the preparation of the remaining grafts was conventional.Results:A total of 30 female patients aged 21-45 years, with an average age of 36 years underwent primary or repaired autogenous costal cartilage rhinoplasty without severe damage to the nasal septum cartilage (at least retention of L-shaped scaffolds larger than 10 mm) were enrolled. All operations were successful, the cartilage scaffold was stable and durable. There were no serious complications such as infection, septal hematoma and cartilage exposure. Postoperative patients were followed up for a duration of 6-24 months. Two out of 30 patients were not satisfied with the postoperative appearance. During the revision surgery, the septal perichondrium was separated, mucosal tissue had grown into the bilateral holes which further verified of the feasibility of drilling. The other patients were satisfactied with the results.Conclusion:Nasal septum extension graft based on drilling technique is beneficial to maintain the long-term stability of cartilage scaffold after comprehensive costal cartilage nasal surgery, which justifies its application in comprehensive nasal plastic surgery.
		                        		
		                        		
		                        		
		                        	
6.Application of nasal septum extension graft drilling technique in rib-based rhinoplasty
Yihao XU ; Jianjun YOU ; Huan WANG ; Ruobing ZHENG ; Lehao WU ; Xulong ZHANG ; Le TIAN ; Junsheng GUO ; Fei FAN
Chinese Journal of Plastic Surgery 2023;39(10):1100-1105
		                        		
		                        			
		                        			Objective:To investigate the clinical application of a cartilage graft fixation method in the context of rib-based rhinoplasty.Methods:The clinical data of patients who underwent costal cartilage based rhinoplasty in the Center of Rhinoplasty and Nasal Reconstruction, Plastic Surgery Hospital, Chinese Academy of Medical Sciences from January 2020 to January 2022 were retrospectively analyzed. A 20 ml syringe needle was used to drill several micropores about 1 mm in diameter on the surface of the cartilage grafts prior to its placement. "septal extension grafts with micropores" were prepared, and the preparation of the remaining grafts was conventional.Results:A total of 30 female patients aged 21-45 years, with an average age of 36 years underwent primary or repaired autogenous costal cartilage rhinoplasty without severe damage to the nasal septum cartilage (at least retention of L-shaped scaffolds larger than 10 mm) were enrolled. All operations were successful, the cartilage scaffold was stable and durable. There were no serious complications such as infection, septal hematoma and cartilage exposure. Postoperative patients were followed up for a duration of 6-24 months. Two out of 30 patients were not satisfied with the postoperative appearance. During the revision surgery, the septal perichondrium was separated, mucosal tissue had grown into the bilateral holes which further verified of the feasibility of drilling. The other patients were satisfactied with the results.Conclusion:Nasal septum extension graft based on drilling technique is beneficial to maintain the long-term stability of cartilage scaffold after comprehensive costal cartilage nasal surgery, which justifies its application in comprehensive nasal plastic surgery.
		                        		
		                        		
		                        		
		                        	
7.Open osteotomy in rhinoplasty
Lehao WU ; Jianjun YOU ; Huan WANG ; Yihao XU ; Ruobing ZHENG ; Xulong ZHANG ; Le TIAN ; Junsheng GUO ; Fei FAN
Chinese Journal of Plastic Surgery 2022;38(3):321-326
		                        		
		                        			
		                        			Objective:To introduce a new method of lateral nasal osteotomy. The key of this method is to fully dissect the tissue around the piriform foramen to achieve nasal osteotomy under direct vision. The osteotomy line in the whole process can be seen, which provides a more reliable control for the degree of osteotomy during the operation, and also makes the postoperative outcome more predictable.Methods:Patients whom were treated with open external osteotomy combined with autologous costal cartilage implantation for augmentation rhinoplasty (with 24 cases of hump nose correction) from October 2018 to October 2020. During the surgery, the whole nasal dorsum was exposed, the superior lateral cartilage was separated from the nasal septum, and the whole process was carried out through the mucosa from the intranasal approach under direct view. The osteotomy line was evaluated at the same time.Results:A total of 33 female patients were retrospectively analyzed. All the operations were successful, and the local swelling occurred after operation. The follow-up time was 3-18 months. After 3 months, the nasal contour and dorsal curves were smooth. Severe infections, major injuries like nasolacrimal duct, lacrimal sac, medial canthus ligament, nerve branches were not noticed in all patients. During the long-term follow-up, two patients exhibited slightly widened nasal bone, but did not require surgical intervention. The shape of nose was greatly improved. Patients were satisfied with the result . The subjective evaluation had "very satisfactory" in 24 cases (72.7%) and "satisfactory" in 9 cases (27.3%). The IGAIS evaluation were also positive: the average score of 33 patients was 1.26±0.34, including 30 cases (90.9%) scoring 1 point and 3 cases (9.1%) scoring 2 points.Conclusions:The open osteotomy technique has better control than that of conventional blind operation. It is more reliable, safer for the surgeons who are new to rhinoplasty and the experienced ones.
		                        		
		                        		
		                        		
		                        	
8.Three-dimensional digital technique-assisted clinical study of subalar change after rhinoplasty with costal cartilage
Le TIAN ; Jianjun YOU ; Lehao WU ; Huan WANG ; Binghang LI ; Yihao XU ; Ruobing ZHENG ; Xulong ZHANG ; Junsheng GUO ; Fei FAN
Chinese Journal of Plastic Surgery 2022;38(7):737-743
		                        		
		                        			
		                        			Objective:To investigate subalar change after rhinoplasty with costal cartilage.Methods:A retrospective study was performed on patients who received cartilaginous rhinoplasty at the Rhinoplasty Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences from May 2019 to August 2020. The facial three-dimensional(3D) and CT scanning data of young female cases was analyzed. Pre-operative and more than 6 months follow-up data was matched. The 3D facial coordinate system was established. Preoperative and postoperative location of the subnasale and the lateral point of nasal-facial junction, the angle between endocanthion-alare and coronal plane and the nasolabial angle were analyzed with paired samples t-test. The volume of subalar area was analyzed with one-sample t-test. Results:Eighteen female cases aged (27.30±4.41) years were enrolled in the study with (10.61±3.53) months follow-up. One case (5.6%) suffered from nostril asymmetry one month after the surgery. The subnasale moved forward from (74.30±1.97) mm to (77.67±2.37) mm [average difference (3.36±0.96) mm]. The lateral point of nasal-facial junction moved forward from (65.51±2.45) mm to (68.05±2.52) mm [average difference (2.53±1.50) mm]. Nasolabial angle was (88.79±11.21) degree preoperatively and (101.37±5.53) degree postoperatively [average difference (12.57±7.57) degree]. Angle between endocanthion-alare and coronal plane increased from (9.01±3.24) degree to (12.73±3.27) degree [average difference (3.72±2.22) degree]. The differences between pre- and post-operative data were statistically significant ( P<0.01). The volume of subalar area increased by (282.59±103.22) mm 3. The difference was statistically significant ( P<0.01). Conclusions:Rhinoplasty with costal cartilage could make subalar area move forward.
		                        		
		                        		
		                        		
		                        	
9.Open osteotomy in rhinoplasty
Lehao WU ; Jianjun YOU ; Huan WANG ; Yihao XU ; Ruobing ZHENG ; Xulong ZHANG ; Le TIAN ; Junsheng GUO ; Fei FAN
Chinese Journal of Plastic Surgery 2022;38(3):321-326
		                        		
		                        			
		                        			Objective:To introduce a new method of lateral nasal osteotomy. The key of this method is to fully dissect the tissue around the piriform foramen to achieve nasal osteotomy under direct vision. The osteotomy line in the whole process can be seen, which provides a more reliable control for the degree of osteotomy during the operation, and also makes the postoperative outcome more predictable.Methods:Patients whom were treated with open external osteotomy combined with autologous costal cartilage implantation for augmentation rhinoplasty (with 24 cases of hump nose correction) from October 2018 to October 2020. During the surgery, the whole nasal dorsum was exposed, the superior lateral cartilage was separated from the nasal septum, and the whole process was carried out through the mucosa from the intranasal approach under direct view. The osteotomy line was evaluated at the same time.Results:A total of 33 female patients were retrospectively analyzed. All the operations were successful, and the local swelling occurred after operation. The follow-up time was 3-18 months. After 3 months, the nasal contour and dorsal curves were smooth. Severe infections, major injuries like nasolacrimal duct, lacrimal sac, medial canthus ligament, nerve branches were not noticed in all patients. During the long-term follow-up, two patients exhibited slightly widened nasal bone, but did not require surgical intervention. The shape of nose was greatly improved. Patients were satisfied with the result . The subjective evaluation had "very satisfactory" in 24 cases (72.7%) and "satisfactory" in 9 cases (27.3%). The IGAIS evaluation were also positive: the average score of 33 patients was 1.26±0.34, including 30 cases (90.9%) scoring 1 point and 3 cases (9.1%) scoring 2 points.Conclusions:The open osteotomy technique has better control than that of conventional blind operation. It is more reliable, safer for the surgeons who are new to rhinoplasty and the experienced ones.
		                        		
		                        		
		                        		
		                        	
10.Three-dimensional digital technique-assisted clinical study of subalar change after rhinoplasty with costal cartilage
Le TIAN ; Jianjun YOU ; Lehao WU ; Huan WANG ; Binghang LI ; Yihao XU ; Ruobing ZHENG ; Xulong ZHANG ; Junsheng GUO ; Fei FAN
Chinese Journal of Plastic Surgery 2022;38(7):737-743
		                        		
		                        			
		                        			Objective:To investigate subalar change after rhinoplasty with costal cartilage.Methods:A retrospective study was performed on patients who received cartilaginous rhinoplasty at the Rhinoplasty Center of Plastic Surgery Hospital, Chinese Academy of Medical Sciences from May 2019 to August 2020. The facial three-dimensional(3D) and CT scanning data of young female cases was analyzed. Pre-operative and more than 6 months follow-up data was matched. The 3D facial coordinate system was established. Preoperative and postoperative location of the subnasale and the lateral point of nasal-facial junction, the angle between endocanthion-alare and coronal plane and the nasolabial angle were analyzed with paired samples t-test. The volume of subalar area was analyzed with one-sample t-test. Results:Eighteen female cases aged (27.30±4.41) years were enrolled in the study with (10.61±3.53) months follow-up. One case (5.6%) suffered from nostril asymmetry one month after the surgery. The subnasale moved forward from (74.30±1.97) mm to (77.67±2.37) mm [average difference (3.36±0.96) mm]. The lateral point of nasal-facial junction moved forward from (65.51±2.45) mm to (68.05±2.52) mm [average difference (2.53±1.50) mm]. Nasolabial angle was (88.79±11.21) degree preoperatively and (101.37±5.53) degree postoperatively [average difference (12.57±7.57) degree]. Angle between endocanthion-alare and coronal plane increased from (9.01±3.24) degree to (12.73±3.27) degree [average difference (3.72±2.22) degree]. The differences between pre- and post-operative data were statistically significant ( P<0.01). The volume of subalar area increased by (282.59±103.22) mm 3. The difference was statistically significant ( P<0.01). Conclusions:Rhinoplasty with costal cartilage could make subalar area move forward.
		                        		
		                        		
		                        		
		                        	
            
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