SummaryHormone-secreting cells within pancreatic islets of Langerhans play important roles in metabolic homeostasis and disease. However, their transcriptional characterization is still incomplete. Here, we sequenced the transcriptomes of thousands of human islet cells from healthy and type 2 diabetic donors. We could define specific genetic programs for each individual endocrine and exocrine cell type, even for rare δ, γ, ε, and stellate cells, and revealed subpopulations of α, β, and acinar cells. Intriguingly, δ cells expressed several important receptors, indicating an unrecognized importance of these cells in integrating paracrine and systemic metabolic signals. Genes previously associated with obesity or diabetes were found to correlate with BMI. Finally, comparing healthy and T2D transcriptomes in a cell-type resolved manner uncovered candidates for future functional studies. Altogether, our analyses demonstrate the utility of the generated single-cell gene expression resource.

Injured tendons require mechanical tension for optimal healing, but it is unclear which genes are upregulated and responsible for this effect. We unloaded one Achilles tendon in rats by Botox injections in the calf muscles. The tendon was then transected and left to heal. We studied mechanical properties of the tendon calluses, as well as mRNA expression, and compared them with loaded controls. Tendon calluses were studied 3, 8, 14, and 21 days after transection. Intact tendons were studied similarly for comparison. Altogether 110 rats were used. The genes were chosen for proteins marking inflammation, growth, extracellular matrix, and tendon specificity. In intact tendons, procollagen III and tenascin-C were more expressed in loaded than unloaded tendons, but none of the other genes was affected. In healing tendons, loading status had small effects on the selected genes. However, TNF-alpha, transforming growth factor-beta1, and procollagens I and III were less expressed in loaded callus tissue at day 3. At day 8 procollagens I and III, lysyl oxidase, and scleraxis had a lower expression in loaded calluses. However, by days 14 and 21, procollagen I, cartilage oligomeric matrix protein, tenascin-C, tenomodulin, and scleraxis were all more expressed in loaded calluses. In healing tendons, the transverse area was larger in loaded samples, but material properties were unaffected, or even impaired. Thus mechanical loading is important for growth of the callus but not its mechanical quality. The main effect of loading during healing might thereby be sought among growth stimulators. In the late phase of healing, tendon-specific genes (scleraxis and tenomodulin) were upregulated with loading, and the healing tissue might to some extent represent a regenerate rather than a scar.

Achilles tendon rupture is a frequent injury with an increasing incidence. Until now, there is no consensus regarding optimal treatment. The aim of this review was to illuminate and summarize randomized controlled trials comparing surgical and non-surgical treatment of Achilles tendon ruptures during the last 10 years. Seven articles were found and they were all acceptable according to international quality assessment guidelines. Primary outcomes were re-ruptures, other complications, and functional outcomes. There was no significant difference in re-ruptures between the two treatments, but a tendency to favoring surgical treatment. Further, one study found an increased risk of soft-tissue-related complications after surgery. Patient satisfaction and time to return to work were significantly different in favor of surgery in one study, and there was also better functional outcome after surgery in some studies. These seven studies indicate that surgical patients have a faster rehabilitation. However, the differences between surgical and non-surgical treatment appear to be subtle and it could mean that rehabilitation is more important, rather than the actual initial treatment. Therefore, further studies will be needed in regard to understanding the interplay between acute surgical or non-surgical treatment, and the rehabilitation regimen for the overall outcome after Achilles tendon ruptures.

Differences in rehabilitation loading pattern in the initial 8 weeks after the repair of an Achilles tendon rupture did not measurably alter the outcome. The time to recover full function after an Achilles tendon rupture is at least 12 months. Registration: NCT02422004 ( ClinicalTrials.gov identifier).

Background: Lysyl oxidase catalyzes collagen cross-link formation, which is essential for mechanically strong collagen fibrils. Results: LOX inhibition stops early mechanical development of tendon constructs and leads to irregularly shaped collagen fibrils. Conclusion: Collagen cross-linking is essential for successful fibrillogenesis and regulates fibril shape. Significance: LOX activity is required in the control of collagen fibril architecture by a mechanism that remains to be explained.

Tendons can function as springs and thereby preserve energy during cyclic loading. They might also have damping properties, which, hypothetically, could reduce risk of microinjuries due to fatigue at sites of local stress concentration within the tendon. At mechanical testing, damping will appear as hysteresis. How is damping influenced by training or disuse? Does training decrease hysteresis, thereby making the tendon a better spring, or increase hysteresis and thus improve damping? Seventy-eight female 10-wk-old Sprague-Dawley rats were randomized to three groups. Two groups had botulinum toxin injected into the calf muscles to unload the left Achilles tendon through muscle paralysis. One of these groups was given doxycycline, as a systemic matrix metalloproteinase inhibitor. The third group served as loaded controls. The Achilles tendons were harvested after 1 or 6 wk for biomechanical testing. An increase with time was seen in tendon dry weight, wet weight, water content, transverse area, length, stiffness, force at failure, and energy uptake in all three groups (P< 0.001 for each parameter). Disuse had no effect on these parameters. Creep was decreased with time in all groups. The only significant effect of disuse was on hysteresis (P = 0.004) and creep (P = 0.007), which both decreased with disuse compared with control, and on modulus, which was increased (P = 0.008). Normalized glycosaminoglycan content was unaffected by time and disuse. No effect of doxycycline was observed. The results suggest that in growing animals, the tendons continue to grow regardless of mechanical loading history, whereas maintenance of damping properties requires mechanical stimulation.

Tendons adapt to changes in mechanical loading, and numerous animal studies show that immobilization of a healing tendon is detrimental to the healing process. The present study addresses whether the effects of a few episodes of mechanical loading are different during different phases of healing. Fifty female rats underwent Achilles tendon transection, and their hind limbs were unloaded by tail suspension on the day after surgery. One group of 10 rats was taken down from suspension to run on a treadmill for 30 min/day, on days 2-5 after transection. They were euthanized on day 8. Another group underwent similar treadmill running on days 8-11 and was euthanized on day 14. Continuously unloaded groups were euthanized on days 8 and 14. Tendon specimens were then evaluated mechanically. The results showed that just four loading episodes increased the strength of the healing tendon. This was evident irrespective of the time point when loading was applied (early or late). The positive effect on early healing was unexpected, considering that the mechanical stimulation was applied during the inflammatory phase, when the calluses were small and fragile. A histological study of additional groups with early loading also showed some increased bleeding in the loaded calluses. Our results indicate that a short episodes of early loading may improve the outcome of tendon healing. This could be of interest to clinical practice. Keywords: early loading; tail-suspension; unloading; mechanical testing; treatmentThe influence of early mobilization on the outcome of Achilles tendon healing has attracted increasing attention. New rehabilitation programs recently tested include early weight bearing 1,2 and the use of orthoses, designed to permit certain ankle movement, and some weight bearing. 3,4 The results from these studies indicate that early mobilization and weight bearing have a positive effect on the healing process. However, clinical research has so far not been able to distinguish between the effects of motion on the readaptation of periarticular tissues and the effects of traction on tendon healing. It is unknown to what extent weight bearing on the injured leg translates into traction forces in the healing Achilles tendon. There is also probably a risk of complications, especially re-rupture, if loading is applied too early. The improved clinical results, however, are in line with previous studies of the effects of mechanical loading in animal models and on tendon cells in vitro. Cyclic loading regimes upregulate both gene expression and production of collagen tendon fibroblasts in vitro. 5,6 This increase in collagen expression can also be seen in intact animal tendons, subjected to increased loading. 7,8 In fact, there are many studies showing that intact tendons adapt to changes in mechanical loading, 9-14 and numerous animal studies show that immobilization of a healing tendon is detrimental for the healing process. [15][16][17][18][19][20] To gain better understanding regarding the effect of mechanical loadin...

Treatment of tendon injuries often involves immobilization. However, immobilization might not prevent mild involuntary isometric muscle contraction. The effect of weak forces on tendon healing is therefore of clinical interest. Studies of tendon healing with various methods for load reduction in rat Achilles tendon models show a consistent reduction in tendon strength by at least half, compared with voluntary cage activity. Unloading was not complete in any of these models, and the healing tendon was therefore still exposed to mild mechanical stimulation. By reducing the forces acting on the tendon even further, we now studied the effects of this mild stimulation. Rat Achilles tendons were transected and allowed to heal spontaneously under four different loading conditions: 1) normal cage activity; 2) calf muscle paralysis induced by botulinum toxin A (Botox); 3) tail suspension; 4) Botox and tail suspension, combined, to eliminate even mild stimulation. Healing was evaluated by mechanical testing after 8 days. Botox alone and suspension alone both reduced tendon callus size (transverse area), thereby impairing its strength compared with normal cage activity. The combination of Botox and suspension did not further reduce tendon callus size but drastically impaired the material properties of the tendon callus compared with each treatment alone. The peak force was only a fifth of that in the normal cage activity group. The results indicate that also the mild loading that occurs with either Botox or suspension alone stimulates tendon healing. This stimulation appears to affect mainly tissue quality, whereas stronger stimulation also increases callus size.