Precisely Tuning the Pore-Wall Surface Composition of Bioceramic Scaffolds Facilitates Angiogenesis and Orbital Bone Defect Repair.

Orbital bone damage (OBD) may result in severe post-traumatic enophthalmos, craniomaxillofacial deformities, vision loss, and intracranial infections. However, it is still a challenge to fabricate advanced biomaterials that can match the individual anatomical structure and enhance OBD repair in situ. Herein, we aimed to develop a selective surface modification strategy on bioceramic scaffolds and evaluated the effects of inorganic or organic functional coating on angiogenesis and osteogenesis, ectopically and orthotopically in OBD models. It was shown that the low thermal bioactive glass (BG) modification or layer-by-layer assembly of a biomimetic hydrogel (Biogel) could readily integrate into the pore wall of the bioceramic scaffolds. The BG and Biogel modification showed appreciable enhancement in the initial compressive strength (∼30-75%) or structural stability in vivo, respectively. BG modification could enhance by nearly 2-fold the vessel ingrowth, and the osteogenic capacity was also accelerated, accompanied with a mild scaffold biodegradation after 3 months. Meanwhile, the Biogel-modified scaffolds showed enhanced osteogenic differentiation and mineralization through calcium and phosphorus retention. The potential mechanism of the enhanced bone repair was elucidated via vascular and osteogenic cell responses in vitro, and the cell tests indicated that the Biogel and BG functional layers were both beneficial for in vitro osteoblastic differentiation and mineralization on bioceramics. Totally, these findings demonstrated that the bioactive ions or biomolecules could significantly improve the angiogenic and osteogenic capabilities of conventional bioceramics, and the integration of inorganic or organic functional coating in the pore wall is a highly flexible material toolbox that can be tailored directly to improve orbital bone defect repair.

Next-generation finely controlled graded porous antibacterial bioceramics for high-efficiency vascularization in orbital reconstruction.

Eyeball loss due to severe ocular trauma, intraocular malignancy or infection often requires surgical treatment called orbital implant reconstruction to rehabilitate the orbital volume and restore the aesthetic appearance. However, it remains a challenge to minimize the postoperative exposure and infection complications due to the inert nature of conventional orbital implants. Herein, we developed a novel Ca-Zn-silicate bioceramic implant with multi-functions to achieve the expected outcomes. The porous hardystonite (Ca2ZnSi2O7) scaffolds with triply periodic minimal surfaces (TPMS)-based pore architecture and graded pore size distribution from center to periphery (from 500 to 800 µm or vice versa) were fabricated through the digital light processing (DLP) technique, and the scaffolds with homogeneous pores (500 or 800 µm) were fabricated as control. The graded porous scaffolds exhibited a controlled bio-dissolving behavior and intermediate mechanical strength in comparison with the homogeneous counterparts, although all of porous implants presented significant antibacterial potential against S. aureus and E. coli. Meanwhile, the pore size-increasing scaffolds indicated more substantial cell adhesion, cell viability and angiogenesis-related gene expression in vitro. Furthermore, the gradually increasing pore feature exhibited a stronger blood vessel infiltrating potential in the dorsal muscle embedding model, and the spherical implants with such pore structure could achieve complete vascularization within 4 weeks in the eyeball enucleation rabbit models. Overall, our results suggested that the novel antibacterial hardystonite bioceramic with graded pore design has excellent potential as a next-generation orbital implant, and the pore topological features offer an opportunity for the improvement of biological performances in orbital reconstruction.

Cathelicidin LL37 Promotes Osteogenic Differentiation in vitro and Bone Regeneration in vivo.

Different types of biomaterials have been used to repair the defect of bony orbit. However, exposure and infections are still critical risks in clinical application. Biomaterials with characteristics of osteogenesis and antibiosis are needed for bone regeneration. In this study, we aimed to characterize the antimicrobial effects of cathelicidin-LL37 and to assess any impacts on osteogenic activity. Furthermore, we attempted to demonstrate the feasibility of LL37 as a potential strategy in the reconstruction of clinical bone defects. Human adipose-derived mesenchyme stem cells (hADSCs) were cultured with different concentrations of LL37 and the optimum concentration for osteogenesis was selected for further in vitro studies. We then evaluated the antibiotic properties of LL37 at the optimum osteogenic concentration. Finally, we estimated the efficiency of a PSeD/hADSCs/LL37 combined scaffold on reconstructing bone defects in the rat calvarial defect model. The osteogenic ability on hADSCs in vitro was shown to be dependent on the concentration of LL37 and reached a peak at 4 µg/ml. The optimum concentration of LL37 showed good antimicrobial properties against Escherichia coli and Staphylococcus anurans. The combination scaffold of PSeD/hADSCs/LL37 showed superior osteogenic properties compared to the PSeD/hADSCs, PSeD, and control groups scaffolds, indicating a strong bone reconstruction effect in the rat calvarial bone defect model. In Conclusion, LL37 was shown to promote osteogenic differentiation in vitro as well as antibacterial properties. The combination of PSeD/hADSCs/LL37 was advantageous in the rat calvarial defect reconstruction model, showing high potential in clinical bone regeneration.

[A brief discussion on theoretical basis and clinical application of "treatment of orofacial diseases by using Hegu(LI4)"].

In traditional Chinese medicine, the well-known saying that "in the treatment of disorders of the orofacial region, Hegu(LI4)is usually taken" is a highly summary of functions of acupoint LI4, the Yuan-primary point of the Large Intestine Meridian of Hand Yangming. By consulting the related literature and viewing from the angle of differentiation of meridians, the present paper expounded the characteristics of location, the attributed meridian and clinical functions of LI4, and summarized its underlying mechanisms in the treatment of some orofacial disorders revealed by experimental researches. In addition, some examples of clinical application of LI4 were listed, including peripheral facial palsy, oral diseases, problems of the nose and headache.

Biomimetic nanofibrous hybrid hydrogel membranes with sustained growth factor release for guided bone regeneration.

Bone regeneration, a complex physiological process, remains a challenge due to the susceptibility to the environment and absence of osteogenic growth factors around the defect region. Although autologous bone grafting is regarded as the gold standard for bone defect treatment, guided bone regeneration membranes in combination with multiple functional growth factors show a striking regeneration effect. Here, a biomimetic nanofibrous hybrid hydrogel composed of bacterial cellulose membranes and alginate/CaCl2 for sustained growth factor delivery was developed. The antibacterial peptide beta-defensin 2 served as an antibacterial, osteogenic, and angiogenic growth factor and was loaded into the aforementioned hydrogel. The mechanical and physical properties of the biomimetic nanofibrous hybrid hydrogel were investigated. Then, the in vitro osteogenic and angiogenic differentiation was confirmed by alkaline phosphatase (ALP) activity, Alizarin Red S staining, qPCR, western blot analysis and tube formation assays. After implantation into a rat calvarial defect model for 12 weeks, nanofibrous hybrid hydrogel membranes could adhere to the defect surface and promote new bone and vessel regeneration.

Effect of Bifunctional ß Defensin 2-Modified Scaffold on Bone Defect Reconstruction.

Bone tissue engineering has emerged as an effective alternative treatment to the problem of bone defect. To repair a bone defect, antibiosis and osteogenesis are two essential aspects of the repair process. By searching the literature and performing exploratory experiments, we found that ß defensin 2 (BD2), with bifunctional properties of antibiosis and osteogenesis, was a feasible alternative for traditional growth factors. The antimicrobial ability of BD2 against Staphylococcus aureus and Escherichia coli was studied by the spread plate and live/dead staining methods (low effective concentration of 20 ng/mL). BD2 was also demonstrated to enhance osteogenesis, with higher messenger RNA (mRNA) and protein expression of the osteogenic markers collagen I (Col1), runt-related transcription factor 2 (Runx2), osteopontin (Opn), and osteocalcin (Ocn) in vitro (1.5-2.5-fold increase compared with the control group in the most effective concentration group), which was consistent with the alkaline phosphatase (ALP) and alizarin red S (ARS) staining results. We implanted poly(sebacoyl diglyceride) (PSeD) combined with BD2 and rat bone tissue-derived mesenchymal stem cells (rBMSCs) under the back skin of rats and found that the inflammatory response was significantly lower with this combination than with the PSeD/rBMSCs scaffold without BD2 and the pure PSeD group and was similar to the control group. Importantly, when assessed in a critical-sized in vivo rat 8 m diameter calvaria defect model, a scaffold we developed combining bifunctional BD2 with porous organic polymer displayed an osteogenic effect that was 160-200% greater than the control group. The in vivo study results revealed a significant osteogenic response and antimicrobial effect and were consistent with the in vitro results. In summary, BD2 displayed a great potential of simultaneously promoting bone regeneration and preventing infection and could provide a viable alternative to traditional growth factors applied in bone defect repair.

[Treatment of Menopausal Syndrome by Combined Electroacupuncture, Acupoint-injection and Fire-needle Therapies].

OBJECTIVE: To observe the clinical efficacy of joint application of electroacupuncture (EA), acupointinjection and fire-needle in the treatment of menopausal syndrome (MS) women. METHODS: One hundred MS patients were randomized into control group and combined treatment group (n=50 patients in each). The combined treatment group was treated by joint application of EA, acupoint-injection and fire-needle stimulation, and the control group treated by EA stimulation alone. EA was applied to Shenshu (BL 23), Xinshu (BL15), Taixi (KI 3), Sanyinjiao (SP 6), Laogong (PC 8), Pishu (BL 20), etc., for 30 min, once daily for 10 days. Point-injection of red ginseng-ophiopogon root injection (0.5 mL) or red-hot needle stimulation was applied to bilateral BL15, BL 20, SP 6 and BL 23 once daily for 10 days and once every other day for 5 times, respectively. The modified Kupperman index scale was used to evaluate symptoms (hot flashes and night sweats: 4 points; paresthesia, insomnia, dyspareunia, irritability and problems of urinary system: 2 point each; depression, dizziness, fatigue, joint pain or muscular pain, headache, palpitation, skin formication: 1 point each) of the perimenopausal syndrome, and 3 months' follow-up survey was conducted after the treatment. Each symptom score was calculated as the product of the symptom severity score (0, no; 1, mild; 2, moderate; 3, severe) multiplied by the points of symptom index. The total Kupperman score was the sum of the scores of all the items. The therapeutic effect was also evaluated according to the "Guiding Principles for Clinical Trials of New Drugs of Chinese Materia Medica" (2002). RESULTS: The Kupperman scores were significantly decreased following the 10 days' treatment and 3 months thereafter in both control and combined treatment groups in comparison with their individual pretreatment (P<0.05), and were obviously lower in the combined treatment group than in the control group after treatment (P<0.05). Of the both 50 patients in the control and combined treatment groups, 22(44%) and 35 (70%) experienced a remarkably improvement, 18(36%) and 12(24%) were effective, and 10(20%) and 3 (6%) ineffective, with the total effective rates being 80% and 94%, respectively. The therapeutic effect of the combined treatment group was notably superior to that of the control group (P<0.05). CONCLUSION: The joint application of electroacupuncture (EA), acupoint-injection and fire needle is significantly superior to that of simple EA in improving the symptoms of patients with menopausal syndrome.

[Clinical Trials of Triple-puncture Stimulation of Zhibian (BL 54), ctc. plus Moxibustion for Benign Prostatic Hyperplasia].

OBJECTIVE: To observe the clinical efficacy of triple-puncture stimulation of Zhibian (BL 54), etc. combined with moxibustion therapy for patients with benign prostatic hyperplasia (BPH). METHODS: Two hundred patients with BPH who met the inclusive criteria were randomized into control (acupuncture) and treatment (acupuncture plus moxibustion) groups (n = 100 cases in each one). Patients of both the control and treatment groups were treated by puncturing the sensitive spot of bilateral Zhibian (BL 54, with triple filiform needles), and other main acupoints as Pang-Shuidao (ST 28, one cun beside ST 28), Pang-Guilai (ST 29, one cun beside ST 29), Shenshu (BL 23), Sanyinjiao (SP 6), Guanyuan (CV 4), Zhongji (GV 7), and some supplemented acupoints selected according to the syndrome identification. In addition, for patients of the treatment group, bird-pecking moxibustion was applied to bilateral Zhibian (BL 54), and the rest acupoints were punctured with filiform needles which were attached with ignited moxa stick-segments. The treatment was conducted for 30 min every time, once every day for 20 times, with 2 days' rest between the two 10 days' treatments. The state of the patients' prostate (micturation state) was evaluated according to the "International prostate symptom score" (I-PSS, 0 - 35 poins, 7 symptom questions) and quality of life (QOL, 0 - 6 points) score before and after the treatment. RESULTS: After the treatment, both l-PSS score and QOL score of the two groups were significantly decreased compared with pre-treatment in the same one group (P<0.01), and the effect of the treatment group was evidently superior to that of the control group in lowering I-PSS score and QOL score (P<0.01). Of the two 100 cases in the control and treatment groups, 23 (23.0%) and 42(42.0%) experienced a marked improvement, 51(51.0%) and 47 (47.0%) were effective, and 26 (26.0%) and 11(11.0%) invalid, with the effective rates being 74.0% and 89.0%, respectively. The therapeutic effect of the treatment group was obviously superior to that of the control group (P<0.01). CONCLUSION: Acupuncture plus moxibustion treatment is effective in improving BPH patients' micturation symptoms, which is superior to the simple acupuncture intervention.

[Clinical observation of lumbar spinal stenosis treated with deep puncture at Jiaji (EX-B 2)].

OBJECTIVE: To explore the clinical efficacy on lumbar spinal stenosis treated with deep puncture at Jiaji (EX-B 2) with round-sharp needle. METHODS: One hundred and fifty cases of lumbar spinal stenosis were divided randomly into a deep puncture at Jiaji (EX-B 2) group (deep puncture group) and a conventional needling group, 75 cases in each one. In deep puncture group, the round-sharp needle was used to puncture Jiaji (EX-B 2) deeply to the nerve root in vertebral canal. Additionally, the conventional acupuncture with filiform needle was applied at the acupoints selected according to the symptoms, such as Shenshu (BL 23), Weizhong (BL 40), Zusanli (ST 36) and Zhibian (BL 54), etc. In conventional needling group, acupuncture with filiform needle was adopted at the acupoints as those in deep puncture group. The clinical symptom scores and efficacies of the patients in two groups were observed after 4 weeks treatment and 3 months of follow-up visit separately. RESULTS: After treatment and in follow-up visit, the clinical symptom scores all increased apparently for the patients in two groups (all P < 0.01). The result in deep puncture group was superior to that in conventional needling group (both P < 0.01). In deep puncture group, the total effective rates were 100.0% (75/75) and 96.0 (72/75) after treatment and in follow-up visit respectively, which were all superior to 92.0% (69/75) and 84.0% (63/75) in conventional needling group (both P < 0.05). CONCLUSION: The deep puncture at Jiaji (EX-B 2) with round-sharp needle achieves superior efficacy as compared with the conventional needling therapy in treatment of lumbar spinal stenosis. The prognosis of it is better and the disease is hardly recurred.

[Clinical observation on three-combination needling method for treatment of primary trigeminal neuralgia].

OBJECTIVE: To probe into a better therapy for primary trigeminal neuralgia. METHODS: Eighty-six cases were randomly divided into an observation group (n = 46) and a control group (n = 40). The observation group were treated with the three-combination needling method, i. e. acupuncture, acupoint-injection and fire-needle therapy, and the control group with acupuncture and acupoint-injection. After treatment of 2 courses, their therapeutic effects were assessed. RESULTS: The total effective rate of 93.5% and the cured rate of 60.9% in the observation group were better than 65.0% and 22.5% in the control group, with significant differences (both P < 0.01). CONCLUSION: The three-combination needling method has obvious clinical therapeutic effect on primary trigeminal neuralgia.

[Clinical study on round-sharp needle therapy combined with massage for prolapse of lumbar intervertebral disk].

OBJECTIVE: To search for a better therapy for prolapse of lumbar intervertebral disk. METHODS: One hundred and sixteen cases were randomly divided into a treatment group treated with round sharp needle plus massage, and a control group with filiform needle plus massage, 58 cases in each group. Changes of symptoms and signs were observed before and after treatment. RESULTS: The effective rate was 98.2% in the treatment group and 82.8% in the control group (P < 0.01). CONCLUSION: Round sharp needle combined with massage has a better therapeutic effect on prolapse of lumbar intervertebral disk.