Wai-Ting Shum

Low-Intensity Pulsed Ultrasound on Tendon Healing A Study of the Effect of Treatment Duration and Treatment Initiation

Background Low-intensity pulsed ultrasound has been reported to be effective in promoting tendon healing. However, its optimal time and duration has not yet been determined. Hypothesis Tendons at different stages of healing may respond differently to low-intensity pulsed ultrasound. In the present study, the timing effects of low-intensity pulsed ultrasound on tendon healing were investigated in a rat model with a patellar tendon graft harvest lesion. Study Design Controlled laboratory study. Methods Sixty Sprague-Dawley rats underwent central third patellar tendon donor site harvest. Low-intensity pulsed ultrasound sonication was then delivered to the injured knees at day 1, 14, or 28 after harvest for 2, 4, or 6 weeks. Tendon samples were harvested at day 14, 28, and 42 after lesion for histological examination and mechanical testing. Results A 2-week session of low-intensity pulsed ultrasound applied from day 1 postlesion (D1-2W) significantly improved the ultimate mechanical strength of the healing tendons from 23.1 ± 8.5 MPa to 36.6 ± 9.0 MPa. Low-intensity pulsed ultrasound did not improve healing when it was given at later stages in D15-2W and D29-2W. When low-intensity pulsed ultrasound treatment was extended from 2 weeks (D1-2W) to 4 weeks (D1-4W) or 6 weeks (D1-6W), the beneficial effects on tendon healing became insignificant. Histological examination showed that low-intensity pulsed ultrasound sonication at late healing stages may disturb remodeling with a poor collagen fiber alignment. Conclusion Low-intensity pulsed ultrasound promoted restoration of mechanical strength and collagen alignment in healing tendons only when applied at early healing stages. Clinical Relevance The present findings indicate that low-intensity pulsed ultrasound may be an effective treatment to reduce tendon donor site morbidity.

The effects of an antiosteoporosis herbal formula containing epimedii herba, ligustri lucidi fructus and psoraleae fructus on density and structure of rat long bones under tail-suspension, and its mechanisms of action.

An innovative anti‐osteoporosis herbal formula containing Epimedii Herba, Ligustri Lucidi Fructus and Psoraleae Fructus (ELP) has been previously shown its bone protecting effects in ovariectomized osteoporotic rats and also in post‐menopausal osteopenic women. This study aimed to investigate the efficacy of ELP against bone loss during physical inactivity or weightlessness. A hindlimb unloading tail‐suspended rat model was used for studying the effects of ELP on bone mineral density (BMD) and bone micro‐architecture. For in vitro mechanistic studies, rat mesenchymal stem cells (MSCs) and mouse macrophage cells (RAW264.7) were used for studying the effects of ELP on osteogenic/adipogenic differentiations and osteoclastogenesis, respectively. Our data illustrated that ELP had a significant preventive effect against bone loss induced by tail‐suspension (TS) at day 28 (p < 0.01) as indicated in the reduction in BMD loss and the preservation of bone micro‐architecture. ELP could significantly promote the osteogenesis and suppress the adipogenesis (p < 0.05) in MSCs. Besides, significant inhibition of osteoclast formation (p < 0.01) was found in ELP‐treated RAW264.7 cells upon receptor activator of nuclear factor kappa‐B ligand induction. Our study presents the first scientific evidence that ELP had a significant preventive effect against bone loss induced by TS through the actions of enhancing osteogenesis, suppressing adipogenesis and osteoclastogenesis. Copyright

In Vivo Low-Intensity Pulsed Ultrasound (LIPUS) Following Tendon Injury Promotes Repair During Granulation but Suppresses Decorin and Biglycan Expression During Remodeling

STUDY DESIGN Bench research, cross-sectional. OBJECTIVE To determine if the effects of low-intensity pulsed ultrasound (LIPUS) on matrix synthesis change at different stages of tendon healing. BACKGROUND LIPUS is effective in promoting tendon healing by stimulation of matrix synthesis. The timing of initiation and duration of LIPUS treatment have been shown to affect its effectiveness to promote tendon healing, suggesting a change of tissue responses to LIPUS stimulation. Understanding how the cellular responses to LIPUS stimulation change during tendon healing is thus important. METHODS In a rat model of patellar tendon donor site injury, a single sonication of LIPUS or mock sonication was delivered to the injured knee of the rats on the fourth, 14th or 28th day postinjury. Tendon samples were harvested at 4 hours and 24 hours after sonication and the mRNA expression of COL1A1, COL3A1, decorin, biglycan, and TGF-beta 1 was analyzed. RESULTS The results showed that a single sonication of LIPUS increased COL1A1 and COL3A1 mRNA in healing patellar tendons when administered on the fourth or 14th day postinjury, but not when administered on the 28th day postinjury. Both decorin and biglycan mRNA were decreased by treatment with LIPUS on the 28th day postinjury. Our results showed that LIPUS enhanced collagen synthesis in vivo only during the granulation phase. Matrix remodeling may be affected by LIPUS with the suppressed expression of decorin and biglycan. CONCLUSION Our findings suggest that LIPUS should be applied during the granulation phase but not during the remodeling phase, to promote tendon healing.

Anti-tumor and anti-osteolysis effects of the metronomic use of zoledronic acid in primary and metastatic breast cancer mouse models

This study aims to determine the effect of metronomic (0.0125 mg/kg twice a week for 4 weeks) zoledronic acid (ZOL) on cancer propagation and osteolysis against both metastatic and primary breast cancer in mice model. From our results, metronomic ZOL resulted in a significant reduction of tumor burden and did not promote lung or liver metastasis. The metronomic ZOL appeared to be more effective than the conventional regimen (0.1 mg/kg once in 4 weeks) in reducing breast cancer tumor burden, and regulating its movement to lung and liver. This dosing schedule of ZOL showed great potential against metastatic breast cancer.

Application of Chinese herbal medicines to revitalize adult stem cells for tissue regeneration.

It has been established in the recent several decades that adult stem cells play a crucial role in tissue renewal and regeneration. Adult stem cells locate in certain organs can differentiate into functional entities such as macrophages and bone cells. Hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) are two of the most important populations of adult stem cells. The application of these stem cells offers a new insight in treating various pathological conditions, through replenishing cells of specific functions by turning on or off the differentiating program within quiescent stem cell niches. Apart from that, they are also capable to travel through the circulation, migrate to injury sites and differentiate to enhance regeneration process. Recently, Chinese medicine (CM) has shown to be potential candidates to activate adult stem cells for tissue regeneration. This review summarizes our own, as well as others’ findings concerning the use of Chinese herbal medicine in the regulation processes of adult stem cells differentiation and their movement in tissue repair and rejuvenation. A number of Chinese herbs are used as therapeutic agents and presumably preventive agents on metabolic disorders. In our opinion, the activation of adult stem cells self-regeneration not only provides a novel way to repair tissue damage, but also reduces the use of targeted drug that adversely altering the normal metabolism of human subjects.

Preclinical evaluations on the efficacy of a topical Chinese herbal formula for swelling control and pain relief.

ETHNOPHARMACOLOGICAL RELEVANCE Patients suffering from musculoskeletal pain and swellings occupy many hospital beds and demand many rehabilitation facilities. Chinese Medicine is offering many alternatives to ameliorate pain and swelling. However, evidence-based scientific publications supporting their efficacy on pain relief are inadequate. The in vitro and in vivo efficacy of a topical use Chinese herbal bath formula (HB) on anti-inflammation and swelling control was studied. MATERIALS AND METHODS The therapeutic mechanisms of HB were studied in vitro via anti-inflammatory and pro-angiogenic assays on RAW264.7 and HUVEC cells, respectively. Fibroblast proliferation was also studied with Hs27 cells. The in vivo angiogenic effect of HB was also studied using zebrafish model, while its efficacy of in vivo anti-Inflammation and swelling control were investigated using rat paw edema model. The affected paw was treated by immersing it in the HB or distilled water as control. The sensation of pain, change in paw thickness and inflammation marker in serum were analyzed. RESULTS In the anti-inflammation assay, HB significantly inhibited nitrite release from RAW264.7 by 47.6% at 800 μg/ml. In the pro-angiogenic assays, it reduced wound area in HUVEC by 8.2% and increased tube formation of HUVEC by 11.5% at 300 μg/ml. HB also stimulated Hs27 proliferation up to 23.5% at 1200 μg/ml. It showed in vivo pro-angiogenic effect by increasing the mean sprout number in the embryos of zebrafish by 2.4 folds. The in vivo therapeutic effects of HB on edema was illustrated by the significant longer thermal withdrawal latency and thinner paw thickness compared with control. After 14 days of treatment, HB also reduced the IL-6 concentration in the serum of rat by 20.9% significantly. CONCLUSIONS This study showed that HB is effective for swelling control and pain relief from edema due to its anti-inflammatory and pro-angiogenic properties.

In vivo study on the pharmacological interactions between a chinese herbal formula ELP and antiresorptive drugs to counteract osteoporosis

Antiresorptive drugs, alendronate and raloxifene, are effective in lowering bone mineral density (BMD) loss in postmenopausal women. However, long-term treatment may be associated with serious side effects. Our research group has recently discovered that a Chinese herbal formula, ELP, could significantly reduce BMD loss in animal and human studies. Therefore, the present study aimed to investigate the potential synergistic bone-protective effects of different herb-drug combinations using ovariectomized rats. To assess the efficacy of different combinations, the total BMD was monitored biweekly in the 8-week course of daily oral treatment. Bone microarchitecture, bone strength, and deoxypyridinoline level were also determined after 8 weeks. From our results, coadministration of ELP and raloxifene increased the total tibial BMD by 5.26% (2.5 mg/kg/day of raloxifene; P = 0.014) and 5.94% (0.25 mg/kg/day of raloxifene; P = 0.026) when compared with the respective dosage groups with raloxifene alone. Similar synergistic effects were also observed in BMD increase at distal femur (0.25 mg/kg/day; P = 0.001) and reduction in urinary deoxypyridinoline crosslink excretion (2.5 and 0.25 mg/kg/day; both P = 0.02). However, such interactions could not be observed in all alendronate-treated groups. Our data provide first evidence that ELP could synergistically enhance the therapeutic effects of raloxifene, so that the clinical dosage of raloxifene could be reduced.

Evaluation of the combined use of metronomic zoledronic acid and Coriolus versicolor in intratibial breast cancer mouse model

ETHNOPHARMACOLOGICAL RELEVANCE Coriolus versicolor (CV) is a mushroom traditionally used for strengthening the immune system and nowadays used as immunomodulatory adjuvant in anticancer therapy. Breast cancer usually metastasizes to the skeleton, interrupts the normal bone remodeling process and causes osteolytic bone lesions. The aims of the present study were to evaluate its herb-drug interaction with metronomic zoledronate in preventing cancer propagation, metastasis and bone destruction. MATERIALS AND METHODS Mice inoculated with human breast cancer cells tagged with a luciferase (MDA-MB-231-TXSA) in tibia were treated with CV aqueous extract, mZOL, or the combination of both for 4 weeks. Alteration of the luciferase signals in tibia, liver and lung were quantified using the IVIS imaging system. The skeletal response was evaluated using micro-computed tomography (micro-CT). In vitro experiments were carried out to confirm the in vivo findings. RESULTS Results showed that combination of CV and mZOL diminished tumor growth without increasing the incidence of lung and liver metastasis in intratibial breast tumor model. The combination therapy also reserved the integrity of bones. In vitro studies demonstrated that combined use of CV and mZOL inhibited cancer cell proliferation and osteoclastogenesis. CONCLUSIONS These findings suggested that combination treatment of CV and mZOL attenuated breast tumor propagation, protected against osteolytic bone lesion without significant metastases. This study provides scientific evidences on the beneficial outcome of using CV together with mZOL in the management of breast cancer and metastasis, which may lead to the development of CV as adjuvant health supplement for the control of breast cancer.

Evaluation of a Topical Herbal Agent for the Promotion of Bone Healing

A topically used Chinese herbal paste, namely, CDNR, was designed to facilitate fracture healing which is usually not addressed in general hospital care. From our in vitro studies, CDNR significantly inhibited the release of nitric oxide from RAW264.7 cells by 51 to 77%. This indicated its anti-inflammatory effect. CDNR also promoted the growth of bone cells by stimulating the proliferation of UMR106 cells up to 18%. It also increased the biomechanical strength of the healing bone in a drill-hole defect rat model by 16.5% significantly. This result revealed its in vivo efficacy on facilitation of bone healing. Furthermore, the detection of the chemical markers of CDNR in the skin and muscle of the treatment area demonstrated its transdermal properties. However, CDNR did not affect the bone turnover markers in serum of the rats. With its anti-inflammatory and bone formation properties, CDNR is found effective in promoting bone healing.

Integrative Approach to Facilitate Fracture Healing: Topical Chinese Herbal Paste with Oral Strontium Ranelate

Strontium ranelate (SrR) is one of the pharmaceutical agents reported to be effective on the promotion of fracture healing. This study aimed to evaluate the integrative effect of the oral SrR with a topical Chinese herbal paste, namely, CDR, on facilitation of bone healing. The in vivo efficacy was evaluated using rats with tibial fracture. They were treated with either CDR topically, or SrR orally, or their combined treatments. The in vivo results illustrated a significant additive effect of CDR on SrR in increasing the yield load of the fractured tibia. The in vitro results showed that neither SrR nor CDR exhibited a cytotoxic effect on UMR106 and bone-marrow stem cell (BMSC), but both of them increased the proliferation of BMSC at low concentrations. The combination of CDR at 200 μg/mL with SrR at 200 or 400 μg/ml also showed an additive effect on increasing the ALP activity of BMSC. Both SrR and CDR alone reduced osteoclast formation, and the effective concentration of SrR to inhibit osteoclastogenesis was reduced in the presence of CDR. This integrative approach by combining oral SrR and topical CDR is effective in promoting fracture healing properly due to their additive effects on proosteogenic and antiosteoclastogenic properties.