Prolotherapy Induces an Inflammatory Response in Human Tenocytes In Vitro

Abstract:

Background: Proliferative therapy, or prolotherapy, is a controversial treatment method for many connective tissue injuries and disorders. It involves the injection of a proliferant, or irritant solution, into the site of injury, which causes small-scale cell death. This therapeutic trauma is theorized to initiate the body’s wound-healing cascade, perhaps leading to tissue repair. The immediate effects of many of these proliferants are poorly characterized, as are the cellular responses to them; here, we sought to evaluate the immediate effects of two common proliferants (dextrose and P2G, a combination of phenol, glucose, and glycerin) on the cellular response of human tenocytes, and begin to explicate the mechanisms with which each proliferant functions.

QUESTIONS/PURPOSES: We asked: What are the effects of treating cultured tenocytes with proliferative treatment agents on their (1) cellular metabolic activity, (2) RNA expression, (3) protein secretion, and (4) cell migration?

Methods: Using human hamstring and Achilles tendon cells, we attempted to answer our research questions. We used a colorimetric metabolic assay to assess the effect of dextrose and P2G proliferant treatment on cell mitochondrial activity compared with nontreated tenocytes. Next, using quantitative PCR, ELISA, and a reporter cell line, we assessed the expression of several key markers involved in tendon development and inflammation. In addition, we used a scratch wound-healing assay to evaluate the effect of proliferant treatment on tenocyte migration.

Results: Results showed that exposure to both solutions led to decreased metabolic activity of tenocytes, with P2G having the more pronounced effect (75% ± 7% versus 95% ± 7% of untreated control cell metabolic levels) (ANOVA; p < 0.01; mean difference, 0.202; 95% CI, 0.052-0.35). Next, gene expression analysis confirmed that treatment led to the upregulation of key proinflammatory markers including interleukin-8 and cyclooxygenase-2 and downregulation of the matrix marker collagen type I. Furthermore, using a reporter cell line for transforming growth factor-β (TGF-β), a prominent antiinflammatory marker, we showed that treatments led to decreased TGF-β bioactivity. Analysis of soluble proteins using ELISA revealed elevated levels of soluble prostaglandin E2 (PGE2), a prominent inducer of inflammation. Finally, both solutions led to decreased cellular migration in the tenocytes.

Conclusion: Taken together, these results suggest that prolotherapy, more so with P2G, may work by decreasing cellular function and eliciting an inflammatory response in tenocytes. Additional studies are needed to confirm the cellular signaling mechanisms involved and the resulting immediate response in vivo.

Read more

Posted in General | Leave a comment

Platelet-Rich Plasma Augmentation for Hip Arthroscopy

Abstract:

Biological augmentation and therapeutics are being increasingly used in musculoskeletal and orthopaedic care. Platelet-rich plasma (PRP) is produced from centrifugation of peripheral blood, a process that concentrates platelets within autologous plasma. The process of PRP preparation is fundamental in controlling the contents, and it influences its therapeutic potential. Platelets contain alpha granules that store and release a variety of growth factors and other proteins that may augment the healing environment; PRP also has the added benefit of promoting postsurgical hemostasis. The purpose of this report was to detail our institutional preparation protocol and method of administration of PRP during hip arthroscopy.

Biological augmentation and therapeutics are being increasingly used in musculoskeletal and orthopaedic care1,2,3 Platelet-rich plasma (PRP) has been used by orthopaedic surgeons and other health practitioners to enhance healing and modulate the environment of tendinopathy, surgical repair of tendons, ligament reconstruction, diffuse arthritis, and focal chondral defects4,5,6.

PRP is produced from centrifugation of peripheral blood, a process that concentrates platelets within autologous plasma.2 Platelets contain alpha granules that store and release a variety of growth factors, such as cytokines and chemokines, as well as other proteins including insulin growth factor 1, transforming growth factor β1, and vascular endothelial growth factor.1 These biological factors can be modulated by the preparation method used to produce the PRP. For example, leukocyte-rich PRP may increase inflammation and catabolic pathways, whereas leukocyte-poor PRP may decrease inflammation and anabolic pathways.7, 8 In addition, PRP can be prepared as a fibrinous product with adhesive hemostatic properties through endogenous or exogenous activation. The PRP-fibrin preparation can enhance endothelial, epithelial, and epidermal regeneration, by stimulating angiogenesis, improving collagen synthesis, and decreasing scarring.9

PRP has been historically described as “a volume of plasma with a platelet count above baseline,”10 but a more recent quantitative definition requires PRP to contain more than 1 million platelets per milliliter of serum or a 5-fold increase from the baseline platelet concentration.11 The preparation of PRP is essential in determining the therapeutic potential for biological augmentation. The purpose of this report was to detail our institutional PRP preparation protocol and method of administration during hip arthroscopy.

Read more

Posted in General | Leave a comment

Single Platelet-Rich Plasma Injection for Early Stage of Osteoarthritis of the Knee

Abstract:

Purpose: The purpose of this study was to determine the safety efficacy and outcomes of platelet-rich plasma (PRP) intra-articular injections for early stages of knee osteoarthritis (OA).

Methods: Twenty-five patients affected by grade I and II knee primary OA according to the Kellgren–Lawrence scale received a single intra-articular PRP injection. Patients were prospectively evaluated for 6 months. Visual analog scale (VAS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and Knee injury and Osteoarthritis Outcome Score (KOOS) scoring scales were used to evaluate clinical outcomes. Wilcoxon signed rank test was used to evaluate significance of improvement of WOMAC, KOOS, and VAS scores.

Results: Twenty-one patients completed 6-months follow-up. The median WOMAC score improved from 29.1 points (range: 17.4–60.4; standard deviation [SD] = 13.0) at baseline to 42.41 (range: 24.3–71.2; SD = 12.5) at final follow-up. Improvements in median KOOS and VAS score have been also found, from 37.49 points and 64.2 mm before injection to 59.71 points and 42.8 mm, respectively. All these improvements were statistically significant (p < 0.05). No adverse reactions have been observed.

Conclusion: Treating knee OA with PRP injection is safe. A single dose of PRP seems to be effective in managing pain and improving quality of life in patients with low-grade knee OA.

Level of Evidence: Level IV, therapeutic case series.

Read more

Posted in General | Tagged , , , , | Leave a comment

Dr Weglein and Dr Garcia teaching at TOBI ( the Orthobiologic institute)

Dr Weglein And Dr Garcia

Posted in General | Leave a comment

TOBI Faculty for 2017

Abstract:

Over the years, TOBI has become best known for world-renowned faculty. TOBI annual events bring together field experts publishing the research in orthopedic biologics and regenerative medicine. This event has enjoyed consistent annual growth, as global thought-leaders return for annual updates and the year’s best networking opportunities to collaborate on projects and advance the field. Newcomers to the field come to learn from the best.

Read more

Posted in General | Leave a comment

Effect of High-Volume Injection, Platelet-Rich Plasma, and Sham Treatment in Chronic Midportion Achilles Tendinopathy: A Randomized Double-Blinded Prospective Study

Abstract:

Background: Injection therapies are often considered alongside exercise for chronic midportion Achilles tendinopathy (AT), although evidence of their efficacy is sparse.

Purpose: To determine whether eccentric training in combination with high-volume injection (HVI) or platelet-rich plasma (PRP) injections improves outcomes in AT.

Study Design: Randomized controlled trial; Level of evidence, 1.

Methods: A total of 60 men (age, 18-59 years) with chronic (>3 months) AT were included and followed for 6 months (n = 57). All participants performed eccentric training combined with either (1) one HVI (steroid, saline, and local anesthetic), (2) four PRP injections each 14 days apart, or (3) placebo (a few drops of saline under the skin). Randomization was stratified for age, function, and symptom severity (Victorian Institute of Sports Assessment–Achilles [VISA-A]). Outcomes included function and symptoms (VISA-A), self-reported tendon pain during activity (visual analog pain scale [VAS]), tendon thickness and intratendinous vascularity (ultrasonographic imaging and Doppler signal), and muscle function (heel-rise test). Outcomes were assessed at baseline and at 6, 12, and 24 weeks of follow-up.

Conclusion: Treatment with HVI or PRP in combination with eccentric training in chronic AT seems more effective in reducing pain, improving activity level, and reducing tendon thickness and intratendinous vascularity than eccentric training alone. HVI may be more effective in improving outcomes of chronic AT than PRP in the short term.

Read more

Posted in General | Leave a comment

Prolotherapy: An effective therapy for Tietze syndrome

Abstract:

Purpose: To compare the short-term effect of prolotherapy and conservative terapy for the Tietze synrome.

Patients And Methods: From 2013 to 2014, twenty-one patients underwent prolotherapy (group 1) and thirteen underwent conservative therapy with analgesics (group 2). A visual analogue score (VAS) was recorded for measurement of pain intensity in all patients before (Pre VAS) and after injection first day (VAS1), first week (VAS2) and fourth week (VAS3). Group 2 were received systemic nonsteroidal anti-inflammatory drug. VAS score was recorded similarly at the same times (Pre VAS, VAS1, VAS2, VAS3), and clinical affects were compared between the two groups.

Results: The mean VAS score (mm) before prolotherapy was 7.10 in patients who received prolotherapy, and 7.14 mm in patients who treated nonsteroidal anti-inflammatory drug. The mean VAS after the fist injection was 2.19 mm and dropped to 1.52 mm after the third injection. The mean VAS after the nonsteroidal anti-inflammatory drug treatment dropped 2.62 mm and during the same scores to 3 weeks later. There was no significant difference between the group 1 and group 2 in the age, sex and comorbidity. Also there was no significant difference between the group 1 and group 2 in clinical and radiological evidence. The prolotherapy group showed a faster recovery, including significantly reduced clinic findings (p: 0.001). Third VAS is significant finding for the prolotherapy group.

Conclusion: Prolotherapy could be performed safely and is a method with a favorable long term treatments for t he Tietze Syndrome. It may be the ideal procedure for patients with drugs side effects and advers events especially for those with limited liver and kidney reserve or significant comorbidities.

Read more

Posted in General | Leave a comment

Houston researchers use stem cells to treat traumatic brain injuries

Abstract:

A Houston research team has uncovered a possible treatment for severe brain injuries that could dramatically improve the outlook for tens of thousands of trauma patients – from those injured on Houston highways, to soldiers wounded on faraway battlefields.

About 275,000 people across the country each year are wheeled into emergency rooms with severe traumatic brain injuries; about a fourth don’t survive. Those who do often suffer permanent disabilities, in part because swelling in the brain in the days and weeks after the injury cuts off blood flow, killing neurons.

In a new clinical trial conducted at Memorial Hermann Hospital, researchers from UTHealth have shown it’s possible to reduce brain inflammation by harvesting stem cells from a trauma patient’s bone marrow and re-infusing them into the bloodstream within 48 hours of injury. The results are promising, said Dr. Charles Cox, who’s been working on the experimental treatment for more than 15 years.

Read more

Posted in General | Leave a comment

Effect of Intraarticular injection of ozone on inflammatory cytokines in knee osteoarthritis

Abstract:

Introduction: Knee Osteoarthrosis is one of the most debilitating diseases. Prolotherapy includes intraarticular injection of various drugs to decrease inflammation. Injection of Intradiscal O2-O3 has reduced pain and disability of patients with low back pain due to prolapsed lumbar disk.

Objective: To compare the effect of intraarticular injection of Ozone and steroids in improvement of clinical and cellular healing of knee osteoarthritis.

Methods:in a randomized clinical trial, 70 patients with knee osteoarthrosis were included in the study. In Ozone group, 5 ml (35 μg/ml) of Ozone and in steroid group 5ml (50 mg) Triamcinolone were injected intraarticular. At 1,2,and 6 month patients were followed for pain scale, disability index and IL-1β and TNF-α serum levels were measured.

Results: At 1 month after injection pain scale and disability index and IL-1β and TNF-α were decreased in both groups. However, at 2 and 6 month pain scale and disability index were significantly lower in Ozone group compare to steroid group (p<0.05). Besides, serum level of IL-1β and TNF-α were also significantly lower at 2 and 6 month in Ozone group compare to steroid group (p<0.05).

Conclusion: Intraarticular Ozone induces significantly longer improvement of pain and disability in knee osteoartherosis compare to steroid injection. In addition, serum inflammatory cytokines are also lower in Ozone group compared to steroid group along with clinical improvements.

Read more

Posted in General | Tagged , , , | Leave a comment

Functional outcomes of platelet rich plasma injections in plantar fascitis

Abstract:

Background: The study was conducted to analyse the clinical outcomes following a single injection of platelet rich plasma, produced using autologous blood by standard double centrifugation technique in patients with plantar fasciitis who have not had relief using conservative methods.

Methods: Between June 2013 and June 2015, 26 consecutive patients (Mean age 45.92) (11 males/15 females) who had symptoms for a minimum of 3 months and had failed conservative management options such as rest, anti-inflammatories and physiotherapy were given a single injection of PRP in the medial calcaneal tubercle.

Results: The outcomes were measured using Visual analogue scale and Foot function index at pre injection, and at 1, 6 and 12 months post injection and results tabulated. All patients reported an improvement in VAS and FFI scores consistently. One superficial infection was encountered.

Conclusions: Single injection of platelet rich plasma improves pain and function in the foot in patients suffering from plantar fasciitis when conservative management has failed. The synergism between various autologous growth factors may play a role in tendon healing.

Read more

Posted in General | Tagged , , | Leave a comment