Asymmetric activation of microglia in the hippocampus drives anxiodepressive consequences of trigeminal neuralgia in rodents.

BACKGROUND AND PURPOSE: Patients suffering from trigeminal neuralgia are often accompanied by anxiety and depression. Microglia-mediated neuroinflammation is involved in the development of neuropathic pain and anxiodepression pathogenesis. Whether and how microglia are involved in trigeminal neuralgia-induced anxiodepression remains unclear. EXPERIMENTAL APPROACH: Unilateral constriction of the infraorbital nerve (CION) was performed to establish trigeminal neuralgia in rat and mouse models. Mechanical allodynia and anxiodepressive-like behaviours were measured. Optogenetic and pharmacological manipulations were employed to investigate the role of hippocampal microglia in anxiety and depression caused by trigeminal neuralgia. KEY RESULTS: Trigeminal neuralgia activated ipsilateral but not contralateral hippocampal microglia, up-regulated ipsilateral hippocampal ATP and interleukin-1ß (IL-1ß) levels, impaired ipsilateral hippocampal long-term potentiation (LTP) and induced anxiodepressive-like behaviours in a time-dependent manner in rodents. Pharmacological or optogenetic inhibition of ipsilateral hippocampal microglia completely blocked trigeminal neuralgia-induced anxiodepressive-like behaviours. Activation of unilateral hippocampal microglia directly elicited an anxiodepressive state and impaired hippocampal LTP. Knockdown of ipsilateral hippocampal P2X7 receptors prevented trigeminal neuralgia-induced microglial activation and anxiodepressive-like behaviours. Furthermore, we demonstrated that microglia-derived IL-1ß mediated microglial activation-induced anxiodepressive-like behaviours and LTP impairment. CONCLUSION AND IMPLICATIONS: These findings suggest that priming of microglia with ATP/P2X7 receptors in the ipsilateral hippocampus drives pain-related anxiodepressive-like behaviours via IL-1ß. An asymmetric role of the bilateral hippocampus in trigeminal neuralgia-induced anxiety and depression was uncovered. The approaches targeting microglia and P2X7 signalling might offer novel therapies for trigeminal neuralgia-related anxiety and depressive disorder.

[Evaluation of digital technology for orthognathic surgery in 25 patients].

PURPOSE: To use three-dimensional reconstruction measurement, preoperative diagnosis, surgical design, surgical simulation, guide plate production, navigation verification and effect evaluation of orthognathic surgery assisted by digital technology, in order to explore more scientific and reasonable programs and procedures of orthognathic surgery. METHODS: Twenty-five patients with congenital dental and maxillofacial deformity were selected as the experimental subjects, craniofacial spiral CT was conducted before surgery and CT data were imported into Mimics 20.0 software to establish a 3D head digital model. The bone landmarks in three-dimensional reconstruction digital model were selected, measured, analyzed and diagnosed, and the design of the surgical plan and the production of the guide plates were performed. Surgical navigation system was used to confirm the maxillary position, verify the bone retention and guide precise bone grinding during operation. Craniofacial spiral CT was conducted 1 week after surgery for postoperative validation of the surgical design protocol. Statistical analysis was performed using SPSS 24.0 software package. RESULTS: All 25 patients were operated according to the digital orthognathic surgery design and procedure.There were no significant differences in X, Y and Z three-dimensional directions in 10 actual landmarks between the postoperative actual head model and the preoperative predictive head model（P>0.05）. CONCLUSIONS: Orthognathic surgery assisted by digital technology has the advantages of precision and minimal invasiveness.

Treatment of oral lichen planus by surgical excision and acellular dermal matrix grafting: Eleven case reports and review of literature.

BACKGROUND: Oral lichen planus (OLP) is a chronic inflammatory disorder, and it can affect normal oral function. The conventional treatments for OLP are not always effective, and relapse easily occurs. Therefore, treatment of OLP is difficult and challenging. In this study, we evaluated over a long period the clinical efficacy of surgical excision and acellular dermal matrix (ADM) grafting in patients with refractory OLP. CASE SUMMARY: Eleven patients with refractory OLP underwent a standardized protocol of surgical excision and ADM grafting. The condition of the area of the grafted wound, the intraoperative maximum mouth opening, pain, and clinical healing were assessed at postoperative follow-up visits. All patients had a flat surgical area with similar mucosal tissue coverage and local scar formation. Patients had no irritation and pain in their mucous membranes when eating acidic and spicy food. All patients' mouth openings returned to normal within 2-6 mo after surgery. During follow-up, none of the patients had recurrence of OLP after surgery. The longest follow-up was 11 yr and the shortest was 6 mo, and none of the patients relapsed during follow-up. CONCLUSION: Surgical excision and ADM grafting could be an effective method to treat refractory OLP.

Distinct Function of Estrogen Receptors in the Rodent Anterior Cingulate Cortex in Pain-related Aversion.

BACKGROUND: Brain-derived estrogen is implicated in pain-related aversion; however, which estrogen receptors mediate this effect remains unclear. This study hypothesized that the different estrogen receptors in the rostral anterior cingulate cortex play distinct roles in pain-related aversion. METHODS: Formalin-induced conditioned place avoidance and place escape/avoidance paradigms were used to evaluate pain-related aversion in rodents. Immunohistochemistry and Western blotting were used to detect estrogen receptor expression. Patch-clamp recordings were used to examine N-methyl-D-aspartate-mediated excitatory postsynaptic currents in rostral anterior cingulate cortex slices. RESULTS: The administration of the estrogen receptor-ß antagonist 4-(2-phenyl-5,7-bis [trifluoromethyl] pyrazolo [1,5-a] pyrimidin-3-yl) phenol (PHTPP) or the G protein-coupled estrogen receptor-1 antagonist (3aS*,4R*,9bR*)-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta [c] quinolone (G15) but not the estrogen receptor-α antagonist 1,3-bis (4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy) phenol]-1H-pyrazole dihydrochloride (MPP) into the rostral anterior cingulate cortex blocked pain-related aversion in rats (avoidance score, mean ± SD: 1,3-bis [4-hydroxyphenyl]-4-methyl-5-(4-[2-piperidinylethoxy] phenol)-1H-pyrazole dihydrochloride (MPP): 47.0 ± 18.9%, 4-(2-phenyl-5,7-bis [trifluoromethyl] pyrazolo [1,5-a] pyrimidin-3-yl) phenol (PHTPP): -7.4 ± 20.6%, and [3aS*,4R*,9bR*]-4-[6-bromo-1,3-benzodioxol-5-yl]-3a,4,5,9b-3H-cyclopenta [c] quinolone (G15): -4.6 ± 17.0% vs. vehicle: 46.5 ± 12.2%; n = 7 to 9; P < 0.0001). Consistently, estrogen receptor-ß knockdown but not estrogen receptor-α knockdown by short-hairpin RNA also inhibited pain-related aversion in mice (avoidance score, mean ± SD: estrogen receptor-α-short-hairpin RNA: 26.0 ± 7.1% and estrogen receptor-ß-short-hairpin RNA: 6.3 ± 13.4% vs. control short-hairpin RNA: 29.1 ± 9.1%; n = 7 to 10; P < 0.0001). Furthermore, the direct administration of the estrogen receptor-ß agonist 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN) or the G protein-coupled estrogen receptor-1 agonist (±)-1-([3aR*,4S*,9bS*]-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta [c]quinolin-8-yl)-ethanone (G1) into the rostral anterior cingulate cortex resulted in conditioned place avoidance (avoidance score, mean ± SD: 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN): 35.3 ± 9.5% and (±)-1-([3aR*,4S*,9bS*]-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta [c]quinolin-8-yl)-ethanone (G1): 43.5 ± 22.8% vs. vehicle: 0.3 ± 14.9%; n = 8; P < 0.0001) but did not affect mechanical or thermal sensitivity. The activation of the estrogen receptor-ß/protein kinase A or G protein-coupled estrogen receptor-1/protein kinase B pathway elicited the long-term potentiation of N-methyl-D-aspartate-mediated excitatory postsynaptic currents. CONCLUSIONS: These findings indicate that estrogen receptor-ß and G protein-coupled estrogen receptor-1 but not estrogen receptor-α in the rostral anterior cingulate cortex contribute to pain-related aversion by modulating N-methyl-D-aspartate receptor-mediated excitatory synaptic transmission.

[Three dimensional study on change ratios of hard and soft tissue after orthognathic surgery].

PURPOSE: To investigate the change ratios of soft and hard tissue after orthognathic surgery three-dimensionally in osseous Class Ⅲ patients, in order to predict postoperative soft tissue three-dimensionally. METHODS: Twenty adult patients were selected as the experimental group, craniofacial spiral CT and three-dimensional (3D) stereophotogrammetry were conducted 2 weeks before surgery and 3 months after surgery. Dolphin imaging software was used to establish 3D image digitizing model and 3D measurement coordinate system. Nineteen soft and hard tissue land marks were selected and matched into 12 pairs. 3D coordinate value of these landmarks were read both before and after surgery and were used for statistical analysis with SPSS 22.0 software package. RESULTS: There was a linear relationship only in Pn/A, Gn'/Gn and Me'/Me in X axis, in Gn'/Gn in Y axis and all pairs of landmarks had a close correlation except UL'/UI and UL/SPr in Z axis. Additionally, the change ratio between soft and hard tissue landmarks of mandible was greater than maxilla. CONCLUSIONS: The changes of soft and hard tissue after orthognathic surgery only lie in Z axis(forward and backward), each pair of matched landmarks exhibits a linear relationship, and the change ratio of mandible is greater than the maxilla.

[Preparation and examination of BMP-2 loaded chitosan nanospheres in vitro].

PURPOSE: To prepare chitosan nanospheres for loading of BMP-2 and to evaluate its size, zeta potential, appearance, degradation and release characteristic in vitro, and then to investigate its feasibility as a carrier for sustained release of BMP-2. METHODS: The BMP-2 loaded chitosan nanospheres were prepared using ionic crosslinking method with tripolyphosphate (TPP) and chitosan. Transmission electron microscope was used to evaluate the morphological properties, and laser particle size analyzer was used to analyze particle size, Zeta potential and distribution. Lysozyme degradation experiment was performed to assess the biodegradation behavior. ELISA assay was used to determine the loading efficiency, encapsulation efficiency and in vitro drug release kinetics. The data was analyzed by SPSS 19.0 software package. RESULTS: The BMP-2 loaded chitosan nanospheres were spherical in shape, smooth on surface and uniform dispersion without aggregation. The mean diameter was 150.85 nm. The dispersion index was 0.37, and zeta potential was +35.42 mV. The average loading efficiency and encapsulation efficiency were (56.83 ± 2.26)% and (68.24 ± 3.83)%, respectively. Release experiment in vitro showed that the releasing property of BMP-2 loaded chitosan nanospheres was consistent with two-phase kinetic regulation and BMP-2 was controlled to release from the chitosan nanospheres over 30 days. CONCLUSIONS: The BMP-2 loaded chitosan nanospheres prepared by ionic crosslinking method are successfully prepared which show a good controlled release property. It provides the basis for further application in bone tissue engineering.

[The change of patients' psychosomatic symptoms after orthognathic surgery].

PURPOSE: To assess the change of patients' psychological status before and after orthognathic surgery. METHODS: The psychological status in 36 patients were assessed before and after orthognathic surgery using the SCL-90 questionnaire. The patients were allowed to evaluate the changes of mastication, facial aesthetics, pronunciation and function of temporomandibular joint subjectively through the questionnaire. The data were analyzed using SPSS13.0 software package. RESULTS: Six factors including obsessive-compulsive symptoms, interpersonal sensitivity, depression, anxiety, phobic anxiety and psychoticism were significantly higher than norm preoperatively. No obvious abnormalities were found 2 years after operation. The self-evaluated quotas by patients after operation were significantly improved compared with those preoperatively. CONCLUSIONS: The patients' psychosomatic symptoms are significantly improved after operation. No significant difference is found compared with ordinary people concerning psychological health.

[Multiple symmetric lipomatosis: clinical analysis of 15 consecutive cases].

PURPOSE: The purpose of this study was to assess the clinical features and available treatment of multiple symmetric lipomatosis (MSL). METHODS: A cohort of 15 cases with MSL registered in the Affiliated Hospital of Qingdao University was reviewed. The etiology, clinical manifestations, treatments and prognosis were also investigated. RESULTS: All the cases were performed sectionalized excisions to cut off the lipoma localized in the neck under general anesthesia. During the follow up period, 13 out of 15 cases were cured after the excision with acceptable appearance and normal local function. Two cases with surgical complications, the one was hematoma, and the other was lymphatic leakage. There were 2 cases with relapses under our follow-up visit. CONCLUSION: Sectionalized lipoma excision under general anesthesia is a relative safe method for MSL.

Ultra-sensitive detection of prion protein with a long range resonance energy transfer strategy.

An ultra-sensitive detection strategy for prion protein is proposed based on the long range resonance energy transfer (LrRET) from quantum dots (QDs) to the surface of gold nanoparticles (AuNPs), in which process energy donor-acceptor separation distance ranges from 9 to 22 nm.

[Study of bone marrow stroma stem cells cell-sheet and endothelial progenitor cells cultured in vitro on allogenic demineralized bone matrix in mongrel].

PURPOSE: To investigate the growth situation of bone marrow stroma stem cells(BMSCs) cell-sheet and endothelial progenitor cells (EPCs) on demineralized bone matrix (DBM) and explore the effect of DBM/BMSCs cell-sheet and DBM/EPCs complex on the construction of functional and vascularized tissue-engineered bone in mongrel. METHODS: BMSC cell-sheet was prepared with cell-sheet engineering approach, EPCs were isolated from canine bone marrow and DBM was prepared from homologous bone. BMSCs cell-sheet and EPCs were inoculated on DBM respectively and observed by inverted phase contrast microscope and scanning electron microscope. The porosity of DBM was measured. RESULTS: The DBM/BMSCs cell-sheet and DBM/EPCs complex were successfully constructed. The BMSCs cell-sheet and EPCs adhered to DBM well and grew rapidly. CONCLUSIONS: The BMSCs cell-sheet and EPCs have good biocompatibility with DBM. The complex of DBM/ BMSCs cell-sheet, DBM/EPCs could provide good conditions to acquire functional and vascularized tissue-engineered bone. Supported by National Natural Science Foundation of China (Grant No.30872896) and Natural Science Foundation of Shandong Province (Grant No.Y2008C77).

Adenosine-aptamer recognition-induced assembly of gold nanorods and a highly sensitive plasmon resonance coupling assay of adenosine in the brain of model SD rat.

In this contribution, we report a molecular recognition between adenosine and its aptamer, which leads to the formation of a four-stranded tetraplex structures (G-quartet) of the aptamer. It is found that the formed G-quartet could induce the side-by-side self-assembly of gold nanorods (AuNRs) owing to the electrostatic interaction between the positive charge of cetyltrimethylammonium bromide (CTAB) on the AuNR surface and the negative charge of the formed G-quartet. Furthermore, the side-by-side self-assembly of AuNRs is characterized by the enhancement of plasmon resonance light scattering (PRLS) signals and the blue-shift of the longitudinal plasmon resonance absorption (LPRA) band owing to the plasmon resonance coupling. Then, based on the enhanced PRLS signals, a simple, highly selective and sensitive detection method for adenosine was developed in the range of 4.0-80.0 nM with the limit of determination of 2.0 nM, which is up to now the best sensitive optical detection method to our knowledge. This method has been successfully applied to the detection of adenosine phosphates in the brains of SD rats, which was in good agreement with a high-performance liquid chromatographic (HPLC) method.

Carbon nanotubes as a low background signal platform for a molecular aptamer beacon on the basis of long-range resonance energy transfer.

Although holding the advantages of both an aptamer and a molecular beacon (MB), a molecular aptamer beacon (MAB) needs complicated and expensive modifications at both of its ends and usually has a high background signal because of the low energy transfer efficiency between the donor and the acceptor. To overcome these shortcomings, in this study, we develop a long-range resonance energy transfer (LrRET) system by separating the donor from the acceptor, wherein only one end of the MAB is fluorescently labeled and acts as the energy donor and multiwalled carbon nanotubes (MWCNTs) are introduced as the energy acceptor. To test the feasibility of the newly designed MAB system, adenosine triphosphate (ATP) has been employed as a proof-of-concept target. It is found that the fluorescence of the designed MAB is completely quenched by MWCNTs, supplying a very low background signal. Then the quenched fluorescence is recovered significantly with the addition of ATP, so that ATP can be detected in the range of 0.8-80 µM with a limit of detection of 0.5 µM (3σ). Compared with the conventional fluorescence resonance energy transfer, the efficiency of LrRET between the dye and MWCNTs is much higher. Since only one end of the MAB needs the modification, the present strategy is simple and cost-effective. Furthermore, the use of MWCNTs can greatly reduce the fluorescence background of the MAB and supply a high sensitivity, showing its generality for detection of a variety of targets.

Aptamer-based silver nanoparticles used for intracellular protein imaging and single nanoparticle spectral analysis.

Aptamer-adapted silver nanoparticles (Apt-AgNPs) were developed as a novel optical probe for simultaneous intracellular protein imaging and single nanoparticle spectral analysis, wherein AgNPs act as an illuminophore and the aptamer as a biomolecule specific recognition unit, respectively. It was found that streptavidin-conjugated and aptamer-functionalized AgNPs show satisfactory biocompatibility and stability in cell culture medium, and thus not only can act as a high contrast imaging agent for both dark-field light scattering microscope and TEM imaging but also can inspire supersensitive single nanoparticle spectra for potential intercellular microenvironment analysis. Further investigations showed that caveolae-related endocytosis is likely a necessary pathway for Apt-AgNPs labeled PrP(c) internalization in human bone marrow neuroblastoma cells (SK-N-SH cells). The integrated capability of Apt-AgNPs to be used as light scattering and TEM imaging agents, along with their potential use for single nanoparticle spectral analysis, makes them a great promise for future biomedical imaging and disease diagnosis.

Cuprous oxide nanospheres as probes for light scattering imaging analysis of live cells and for conformation identification of proteins.

A facile solution-phase synthesis route of highly uniform Cu(2)O nanospheres (Cu(2)O NPs) with the size of 57.7+/-4.7nm was developed, and then the nanoparticles were applied to live cell imaging under a common dark-field microscope. Starting from copper(II) salts, the synthesis of Cu(2)O NPs was made in the presence of cetyltrimethylammonium bromide (CTAB) by reducing the copper(II) with sodium borohydride (NaBH(4)) in aqueous medium and by aging process in the air. Monitoring of morphology evolution process of Cu(2)O NPs with scanning electron microscopy (SEM) and measuring of the UV-visible spectra showed that the synthesis of Cu(2)O NPs follows the reduction-oxidation coupled process of Cu(2+) into Cu(0) species at first and then the resulted Cu(0) species into Cu(2)O NPs in the air. Light scattering (LS) features have been measured with a common spectrofluorometer and a common dark-field microscope, and it was found that the as-prepared Cu(2)O NPs display strong blue scattering light and can be applied for cell imaging. If incubated with human bone marrow neuroblastoma, transferrin-conjugated Cu(2)O NPs can get into the cells and show strong pure blue light in cytoplasm. Further investigations showed that the Cu(2)O NPs could be applied for probes for conformation of proteins.

Effect of platelet-rich plasma and latissimus dorsi muscle flap on osteogenesis and vascularization of tissue-engineered bone in dogs.

PURPOSE: The present study evaluated the effects of platelet-rich plasma (PRP) and the latissimus dorsi muscle flap on osteogenesis and vascularization of tissue-engineered bone. MATERIALS AND METHODS: Bone marrow stromal cells (BMSCs) were subcultured, and PRP was obtained from the same dogs. Demineralized bone matrix (DBM) was prepared from homologous bone. The complexes of DBM/BMSCs/PRP were implanted into areas A and B on the left side of the dogs' backs; complexes of DBM/BMSCs without PRP were implanted in areas C and D on the right side of the same dog. The implants in areas A and C were wrapped with a latissimus dorsi muscle flap, and the implants in areas B and D were wrapped with inferior fascia. At 4, 8, and 12 weeks later, the implants were removed for evaluation. RESULTS: The radiographic evaluation, descriptive histologic analysis, and histologic quantitative analysis showed that the PRP/BMSCs/DBM complex was better than the BMSCs/DBM complex in both vascularization and osteogenesis of the ectopic tissue-engineered bones, and the complex wrapped with the latissimus dorsi muscle flap was better than that packed with superficial fascia without blood vessels. CONCLUSIONS: The PRP and blood vessels in the latissimus dorsi muscle could cooperatively promote osteogenesis and vascularization in tissue-engineered bone.

[Effect of platelet rich plasma on vascularization of tissue-engineered bone].

OBJECTIVE: To study the effects of platelet rich plasma (PRP) on vascularization of tissue-engineered bone. METHODS: Bone marrow stromal cell (BMSC) were isolated from iliac bone of dogs. PRP was obtained from the same dog and demineralized bone matrix (DBM) was prepared from homologous bone. Twelve dogs were divided into three groups and the back of each dog was divided into four areas. The DBM- BMSC- PRP was implanted in the area A and B; the DBM-BMSC without PRP was implanted in the area C and D. The implants in the areas A and C were wrapped using myo-fascia with blood vessel of latissimus dorsi. The implants in the area B and D were wrapped using superficial fascia of the back without blood vessel. The implants were taken out 4, 8 and 12 weeks later for examination. RESULTS: The degree of calcification and blood vessel formation of the implants was A > B > C > D. CONCLUSIONS: Both PRP and vessels of latissimus dorsi muscle could promote calcification and vascularization in tissue-engineered bone, when used separately or in combination.

[Effect of platelet-rich plasma and latissimus dorsi myofascia with blood vessel on vascularization of tissue engineered bone in dogs].

OBJECTIVE: To study the effect of platelet-rich plasma (PRP) and latissimus dorsi myofascia with blood vessel on vascularization of tissue engineered bone in dogs. METHODS: Bone marrow stromal cells (BMSCs) were isolated from iliac bone of dogs. PRP was obtained from the same dog. And demineralized bone matrix (DBM) were prepared from homologuous bone. ABCD 4 areas were divided on the back of dog. PRP/BMSCs/DBM was implanted around the vessels of lattisimus dorsi muscle in the A. PRP/BMSCs/DBM wrapped by superficial fascia in the B. BMSCs/DBM was implanted around vessels of lattisimus dorsi muscle in the C. BMSCs/DBM wrapped by superficial fascia in the D area of the same dog. 4, 8, 12 weeks after implantation, gross specimen and histology examination were made. RESULTS: Osteogenesis and blood vessel formation results were A>B>C>D area. CONCLUSION: The results suggested that the PRP and latissimus dorsi myofascia with blood vessels could promote calcification and vascularization in tissue-engineered bone.

[Study of MSCs in vitro cultured on demineralized bone matrix of mongrel].

PURPOSE: To manufacture demineralized bone matrix(DBM) of mongrel and to explore the feasibility of DBM as scaffold of bone tissue engineering. METHODS: Thigh bones of mongrel were degreased, demineralized, deproteined, freezed, dried and sterilized to form DBM. Mesenchymal stem cells(MSCs) were seeded onto the scaffold and their growth were examined by inverted phase contrast microscope and scanning electron microscope. RESULTS: DBM had a three-dimensional mesh structure.The mean pore diameter of DBM was (254.39+/-88.71)microm and the pore rate was about 70%.MSCs could adhere to the surface and inner walls of DBM, proliferated well and secreted a large amount of extracellular matrix. CONCLUSION: DBM has satisfactory biocompatibility.