1.Effect of different concentrations of fetal bovine serum on bone marrow mesenchymal stem cell purity and cycle
Acta Universitatis Medicinalis Anhui 2014;(6):742-747
Objective To compare the influence of three kinds of complete media with 0. 10 ,0. 15 ,0. 20 fetal bo-vine serum( FBS) on purity and cycle of rat bone marrow mesenchymal stem cells ( BMSCs) cultured in vitro and to seek suitable FBS concentration for the cultivation of the stem cells. Methods SD rats were executed by cervical dislocation method and used whole bone marrow adherence to isolate rat BMSCs. Experiment was divided into A, B,C,3 groups. Compared the expression of CD45, CD29, CD90, CD44 in the three groups in 2,3,4,5 passages ( P2 , P3 , P4 , P5 );the cells of P3 in group A were digested and cultured in three different concentration of com-plete culture media for four days, measuring cell cycle in 24,48,72,96 h by flow cytometry instrument. BMSCs of P3 were collected and inoculated to 6 pieces of 96-well plates, then vaccinated with complete media with 0. 10, 0. 15,0. 20 FBS in every plate, one culture plate was taken out for optical density(OD) meaturement every day with CCK-8. Results CD45 was negative, CD29, CD90, CD44 were positive. The difference of BMSCs surface markers cultured in the three kinds of complete media was bigger in the first two passages, but the difference was less in P3 , P4 , all could obtain pure BMSCs in P4 relatively;according to the results of the cell cycle at the same time, G0/G1 phase:with the increase of concentration of fetal bovine, G0/G1 phase reduced and had no diference in A, B, C groups;G2/M phase:there was difference between them after 24 h(P<0. 05,F=12. 412), but with the extension of time, the differences disappeared;S phase:there was no difference in the three groups;S+G2/M phase increased with the concentration of FBS. According to the result of cell vitality, the OD of ABC three groups increased in turn in 24 h, and there was difference(P<0. 05,F=5. 002), but with the extension of time, there was no obvious difference between them. Conclusion The three kinds of culture media in P4 can obtain pure BM-SCs, cell cycle and vitality show three complete media can promote the growth of BMSCs. There is no difference between them. Culture media with 0. 10 FBS can satisfy the isolation and amplification of BMSCs, in order to ob-tain pure BMSCs in the short term, using culture media with 0. 15 and 0. 10 FBS in the primary culture and subcul-ture respectively.
2.Fabrication of hydroxyapatite-chitosan composite microspheres and their biological effect study in vitro
Acta Universitatis Medicinalis Anhui 2015;(11):1552-1555
Objective To investigate the influence of HAp-CS composite microsphere scaffold on the in vitro cell behaviors of mesenchymal cells and evaluate its potential application for bone tissue engineering.Methods Nano-hydroxyap-atite (HAp)and chitosan (CS)composites solution were assembled into microsphere scaffold through microfluidic and observed by inverted microscope.Rat bone marrow mesenchymal stem cells were co-cultured in vitro with the microspheres for calculating the adhesion rate for the first 6h.Proliferation rate was measured by cell counting in the next 1,3,6,9 d,respectively,and GraphPad Prism 6 software was used for statistical analysis.The morphology of BMSCs on the surface of HAp-CS composite microsphere was observed by scanning electron micros-copy (SEM)and confocal scanning microscopy.The cells and HAp-CS microspheres were filled into a disc mold and co-cultured for 14 ~21 d to observe the morphology.Results HAp-CS microspheres were observed to be round and with uniform size by microscope.The adhesion rate of BMSCs reached 80% after cultured for 6 h,and proliferation rate reached the highest value when cultured for 6 d.SEM observations showed that BMSCs adhered compactly to the surface of the microspheres,and the microspheres could be connected together through BMSCs.Af-ter co-culturing BMSCs with microspheres for 14 ~21 d,a complete tissue constructs could be formed.Conclusion HAp-CS microspheres are proved to be good scaffolds for promoting BMSCs adhesion and proliferation.Large a-mount of extracellular matrix can be formed to connect microspheres after co-cultured for a certain time,which paves the way for HAp-CS microspheres to be applied for bone regeneration in animal experiments.
3.Measurement and analysis anatomy factors of maxillary canine fossa related to implanting technology by cone-beam CT
Acta Universitatis Medicinalis Anhui 2017;52(6):925-929
Cone-beam computed tomography(CBCT) was used to analyze the anatomic morphology of maxillary canine fossa of different sagittal skeletal patterns patients;implants of various dimensions were virtually placed into the maxillary premolars region.The relationships among maxillary canine fossa and implant were analyzed.The depth of the maxillary canine fossa was(4.33±0.73),(3.77±0.58),(5.18±0.93)mm in the 1st premolar region of class Ⅰ,Ⅱ and Ⅲ respectively.The depth of the maxillary canine fossa was(3.20±0.63),(2.81±0.58),(3.90±0.79)mm in the 2nd premolar region of class Ⅰ,Ⅱ and Ⅲ respectively.There were no significant statistical differences in genders,sides,and loss of tooth.And there were significant statistical differences in three classes(P<0.05).The length of the implants was (10.30±1.70)mm(d=3.3 mm) and (8.77±1.58)mm(d=4.1 mm) in the 1st premolar region and (8.09±1.51)mm(d=3.3 mm),(6.69±1.35)mm(d=4.1 mm) in the 2nd premolar region in patients with teeth and buccal perforation;the length of the implants was (8.98±1.54)mm(d=3.3 mm) and (7.67±1.52)mm(d=4.1 mm) in the 1st premolar region and(7.09±1.59)mm(d=3.3 mm) and (5.79±1.34)mm(d=4.1 mm) in the 2nd premolar region in patients with tooth loss,respectively.There were significant statistical differences in loss of tooth(P<0.05).Analyzing of spatial relationships among maxillary canine fossa and implants of different sagittal skeletal patterns patients especially class Ⅲ patients,by using CBCT has important significance in guiding dental implants.
4.Application of Endopore implant in inadequate alveolar jaw bone
Chinese Journal of Tissue Engineering Research 2007;0(36):-
From March 2004 to June 2007, the effect of the 32 Endopore implants which were placed in 25 patients with the inadequate alveolar jaw bone treated in the Implant Center of Department of Oral and Maxillofacial Surgery, the Affiliated Stomatological Hospital of Anhui Medical University were retrospectively analyzed. All patients were treated with Endopore implants of 5 mm ? 7 mm without any bone grafts simultaneously. Second-stage restoration was conducted 3-6 months later. Regular follow-up was between 1 and 4 years. All of the 32 implants gained osseointegration and were restored. During the follow-up, no implant loosed or lost. No inflammation was observed in the soft tissue around the implants. All the patients were satisfied with the final prostheses. With the unique root surface of porous microscopic titanium particle and the shorter design, Endopore implants can be used in the posterior area of inadequate alveolar jaw bone. They could improve the indication of the dental implant.
5.Clinical study of staged surgery technique in repairing bilateral mixed cleft lip
Xianyu ZHENG ; Jiguang CHENG ; Jiacai HE
Acta Universitatis Medicinalis Anhui 2016;51(7):1074-1076
8 cases of children with bilateral mixed cleft lip were repaired by two staged surgeries .Results showed that the incision of the two staged surgeries was healed in phase Ⅰ.After the second -staged surgery, the height of bilateral lips was almost symmetrical, the bilateral nostrils had good symmetry and the red lip had natural form .The research shows that the operational effect of the repair for bilateral mixed cleft lip is good by two staged surgeries . The method can be accepted by the children 's family, it is worth promoting.
6.Effects of cellular microenvironment on the efficacy of biomacromolecular therapies
Mohan WANG ; Duohong ZOU ; Jiacai HE
International Journal of Biomedical Engineering 2014;37(4):247-250
Currently,with the rapid development of drug delivery technologies,more and more efforts are taken into the efficient therapies for various diseases by delivering biologically-active macromolecules into the target cells directly.Although a certain number of positive treatment results were obtained from the therapies by using the biomacromolecules to cure some diseases,the microenvironment around the target cell still has a great influence on the final treatment effect.Since many diseases and injuries interfere the normal architecture of the extracellular matrix (ECM),the cell adhesion to ECM and the subsequent cellular activities,the normal microenvironment of the cell plays a critical role in maintaining body balance,tissue regeneration and repair.Given these points,this paper reviews the effects of the cellular microenvironment constructed by ECM on the efficacy of bioactive macromolecules,and provides a theoretical basis for future drug design and synthesis of pharmaceuticals.
7.Clinical application of VIP-CT flap with GBR technique in dental implantation of the maxillary anterior region
Xinxiu DUAN ; Xin LIU ; Jiacai HE
Acta Universitatis Medicinalis Anhui 2015;(4):549-551
This article presented a series of cases using vascularized interpositional periosteal-connective tissue ( VIP-CT) flap with guide bone regeneration ( GBR) in peri-implant soft and hard tissue reconstuction at the esthet-ic zone of maxillary. Fifteen cases with bone and soft tissue defects underwent VIP-CT flap with GBR in the implant treatment. And the attached gingiva width was evaluated before treatment and six months and eighteen months after the operation. The width of attached gingival of six months and eighteen months after surgery was significantly dif-ferent from the preoperative value (P<0. 05). However, no statistically significant difference could be found at six months and eighteen months postoperative. The application of VIP-CT flap could increase the width of attached gin-giva around implants and the short-term effects were stable and favorable.
8.Preparation of sodium alginate-nanohydroxyapatite composite material for bone repair and its biocompatibility.
Yanmei WANG ; Jiacai HE ; Quanli LI ; Jijia SHEN
West China Journal of Stomatology 2014;32(1):27-31
OBJECTIVETo prepare sodium alginate-nanohydroxyapatite composite material and to explore its feasibility as a bone repair material.
METHODSSodium alginate-nanohydroxyapatite composite material was prepared using chemical cross-linking and freeze-drying technology. The composite was characterized by X-ray diffraction (XRD) and scanning electron microscope (SEM) and its porosity was measured by liquid displacement method. The fifth passage of bone marrow stromal stem cells (BMSCs) were incubated on the composite material and then growth was observed by inverted microscope and SEM. BMSCs were cultured with liquid extracts of the material, methyl thiazolyl tetrazolium (MTT) assay was used to calculate the relative growth rate (RGR) on 1, 3, 5 d and to evaluate the cytotoxicity. Fresh dog blood was added into the liquid extracts to conduct hemolysis test, the spectrophotometer was used to determine the optical density (OD) and to calculate the hemolysis rate.
RESULTSSodium alginate-nanohydroxyapatite composite material displayed porosity, the porous pore rate was (88.6 +/- 4.5)%. BMSCs showed full stretching and vigorous growth under inverted microscope and SEM. BMSCs cultured with liquid extracts of the material had good activities. The toxicity of composite material was graded as 1. Hemolysis test results showed that the hemolysis rate of the composite material was 1.28%, thus meeting the requirement of medical biomaterials.
CONCLUSIONThe composite material fabricated in this study has high porosity and good biocompatibility.
Alginates ; Biocompatible Materials ; Cells, Cultured ; Glucuronic Acid ; Hexuronic Acids ; Humans ; Mesenchymal Stromal Cells ; Porosity ; Tissue Engineering ; Tissue Scaffolds
9.Investigation and analysis on the employment issues of postgraduates majoring in clinical science of stomatology
Duohong ZOU ; Siheng ZHU ; Kai ZHANG ; Wei HAN ; Jiacai HE ; Jian ZHOU
Chinese Journal of Medical Education Research 2012;11(4):371-374
Objective To investigate the preference of the postgraduates in school for work and their psychological changes based on the analysis of the employment status of postgraduates majoring in clinical science of stomatology in Anhui medical university in recent five years in an aim to provide preemployment counseling.Methods The employment status of postgraduates majoring in clinical science of stomatology in affiiated stomatological hospital of Anhui medical university in recent five years (2006 -2010) was summed up.Questionnaire of employment preference and the psychological pressure was conducted in 66 postgraduates majoring in clinical science of stomatology (2008 -2010 grade).Results The employment rate of provincial first-class hospitals was obviously decreased in the recent five years while that of municipal first-class hospitals was increased gradually.The number of postgraduates willing to work in the municipal first-class hospitals was reduced while that willing to future their study and work in the non first-class hospitals was increased.Psychological pressure was univcrsally existed in students and did not get released.Conelusion As the employment situation for postgraduates majoring in clinical science of stomatology changes,the preference for work and psychological pressure of the postgraduates also undergo corresponding changes.Targeted measures can be formulated to work out the problem based on the understanding of these changes.Meanwhile,adjustment should be made in the training methods and employment guidance for postgraduates majoring in clinieal science of stomatology.
10.Clinical effect of platelet-rich fibrin combined with bone substitute in alveolar ridge preservation
ZHANG Yun ; WANG Yanmei ; HE Jiacai
Journal of Prevention and Treatment for Stomatological Diseases 2020;28(6):361-366
Objective:
To study the clinical outcomes of implanting platelet rich fibrinogen (PRF) mixed with Bio-Oss® in the extraction socket for alveolar ridge preservation and to provide evidence for clinical application.
Methods:
Thirty-six patients who underwent alveolar ridge preservation were enrolled. Thirty-six extraction sites were divided into two groups: PRF mixed with Bio-Oss® group (test group) and Bio-Oss® alone (control group). Bone dimensional changes in height and width were measured by CBCT before and 6 months after surgery, and early soft tissue healing and postoperative pain sensation were evaluated clinically 1 week after surgery.
Results :
There was no significant difference in the alveolar bone height (-1.48 ± 0.40) mm between the test group and the control group. The difference in the alveolar bone width between the test group (-1.09 ± 0.42) mm and the control group (-1.35 ± 0.22) mm was statistically significant (z=-2.63, P=0.01). The postoperative pain score of the test group was 2.39 ± 1.20, and that of the control group was 3.39 ± 1.65, the difference was statistically significant (t=-2.083, P=0.045). There was no significant difference in soft tissue healing between the test group and the control group.
Conclusion
The use of PRF mixed with Bio-Oss ®in the alveolar ridge preservation procedure can reduce alveolar bone absorption and postoperative pain.