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Hammerman, Malin
Publications (5 of 5) Show all publications
Hammerman, M. (2018). Tendon Healing: Mechanical Loading, Microdamage and Gene Expression. (Doctoral dissertation). Linköping: Linköping University Electronic Press
Open this publication in new window or tab >>Tendon Healing: Mechanical Loading, Microdamage and Gene Expression
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Mechanical loading and the inflammatory response during tendon healing might be important for the healing process. Mechanical loading can improve the healing tendon but the mechanism is not fully understood. The aim of this thesis was to further clarify the effect of mechanical loading on tendon healing and how mechanical loading affects the inflammatory response during the healing process.

We used a rat Achilles tendon model to study healing. The rats were exposed to different degrees of loading by unloading methods such as paralysis of the calf muscles with Botox, tail suspension, and an orthosis (a boot). Full loading was achieved by free cage activity or treadmill walking. Microdamage in tendons, unloaded with Botox, was also investigated by needling. The healing tendons were evaluated in a materials testing machine (to analyze the mechanical properties), by gene expression analysis (microarray and PCR), or histology.

Our results show that moderate loading (unloading with Botox) improves the mechanical properties of healing tendons compared to minimal loading (unloading with Botox in combination with tail suspension or a boot), especially the material properties. In accordance to these findings, expression of extracellular matrix genes were also increased by moderate compared to minimal loading.

Full loading improved all mechanical properties and the expression of extracellular matrix genes was further increased compared to moderate loading. However, structural properties, such as the strength and the size of the healing tendon, were more affected by full loading. Full loading also affected the expression of inflammation-related genes during the early healing phase, 3 and 5 days after tendon injury, and increased the number of immune cells in the healing tendon tissue. Also microdamage of the healing tendon (detected by blood leakage) was increased by full loading compared to moderate loading during the early healing phase.

Induced microdamage by repeated needling in the healing tendon tissue increased the structural properties of the healing tendon. The gene expression after needling was similar to the gene expression after full loading.

The improvement of mechanical properties by loading in healing tendons was decreased by an anti-inflammatory drug called parecoxib, which decreases the production of prostaglandins by inhibiting COX-2 activity. The effect of parecoxib was reduced when loading was reduced but we could not confirm that the effect of parecoxib was related to the degree of loading. However, parecoxib abolished the stimulatory effect of microdamage.

In conclusion, these studies show that moderate loading improves the quality of the healing tendon whereas full loading also increases the quantity of the healing tendon tissue. Full loading creates microdamage and increases inflammation during the early healing phase. The strong effect of full loading on the structural properties might be due to microdamage. Indeed, the anti-inflammatory drug parecoxib seems to impair mechanical stimulation of healing tendons by reducing the response to microdamage.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2018. p. 28
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 1609
National Category
Biomaterials Science
Identifiers
urn:nbn:se:liu:diva-145281 (URN)10.3384/diss.diva-145281 (DOI)9789176853610 (ISBN)
Public defence
2018-03-26, Belladonna, Campus US, Linköping, 13:00 (English)
Opponent
Supervisors
Available from: 2018-02-20 Created: 2018-02-20 Last updated: 2018-02-28Bibliographically approved
Hammerman, M., Blomgran, P., Ramstedt, S. & Aspenberg, P. (2015). COX-2 inhibition impairs mechanical stimulation of early tendon healing in rats by reducing the response to microdamage. Journal of applied physiology, 119(5), 534-540
Open this publication in new window or tab >>COX-2 inhibition impairs mechanical stimulation of early tendon healing in rats by reducing the response to microdamage
2015 (English)In: Journal of applied physiology, ISSN 8750-7587, E-ISSN 1522-1601, Vol. 119, no 5, p. 534-540Article in journal (Refereed) Published
Abstract [en]

Early tendon healing can be stimulated by mechanical loading and inhibited by cyclooxygenase (COX) inhibitors (nonsteroidal anti-inflammatory drugs). Therefore, we investigated if impairment of tendon healing by a COX-2 inhibitor (parecoxib) is related to loading. Because loading might infer microdamage, which also stimulates healing, we also investigated if this effect is inhibited by parecoxib. The Achilles tendon was transected in 114 rats. Three degrees of loading were used: full loading, partial unloading, and unloading (no unloading, Botox injections in the plantar flexor muscles, or Botox in combination with tail suspension). For each loading condition, the rats received either parecoxib or saline. In a second experiment, rats were unloaded with Botox, and the tendon was subjected to microdamage by needling combined with either saline or parecoxib. Mechanical testing day 7 showed that there was a significant interaction between loading and parecoxib for peak force at failure (P less than 0.01). However, logarithmic values showed no significant interaction, meaning that we could not exclude that the inhibitory effect of parecoxib was proportionate to the degree of loading. Microbleeding was common in the healing tissue, suggesting that loading caused microdamage. Needling increased peak force at failure (P less than 0.01), and this effect of microdamage was almost abolished by parecoxib (P less than 0.01). Taken together, this suggests that COX-2 inhibition impairs the positive effects of mechanical loading during tendon healing, mainly by reducing the response to microdamage.

Place, publisher, year, edition, pages
AMER PHYSIOLOGICAL SOC, 2015
Keywords
tendon healing; COX-2; NSAIDs; mechanical stimulation; microdamage
National Category
Physiology Pharmacology and Toxicology
Identifiers
urn:nbn:se:liu:diva-122063 (URN)10.1152/japplphysiol.00239.2015 (DOI)000360694300013 ()26159755 (PubMedID)
Note

Funding Agencies|Swedish Research Council [K2013-52X-02031-47-5]; Swedish National Centre for Research in Sports; King Gustaf V and Queen Victoria Free Mason Foundation

Available from: 2015-12-18 Created: 2015-10-19 Last updated: 2019-02-11
Hammerman, M., Aspenberg, P. & Eliasson, P. (2014). Microtrauma stimulates rat Achilles tendon healing via an early gene expression pattern similar to mechanical loading. Journal of applied physiology, 116(1), 54-60
Open this publication in new window or tab >>Microtrauma stimulates rat Achilles tendon healing via an early gene expression pattern similar to mechanical loading
2014 (English)In: Journal of applied physiology, ISSN 8750-7587, E-ISSN 1522-1601, Vol. 116, no 1, p. 54-60Article in journal (Refereed) Published
Abstract [en]

Mechanical loading increases the strength of healing tendons, but also induces small localized bleedings. Therefore, it is unclear if increased strength after loading is a response to mechanotransduction or microtrauma. We have previously found only five genes to be up-regulated 15 min after a single loading episode, of them four were transcription factors. These genes are followed by hundreds of genes after 3 h, many of them involved in inflammation. We now compared healing in mechanically unloaded tendons with or without added microtrauma induced by needling of the healing tissue. Nineteen rats received Botox into the calf muscle to reduce loading, and the Achilles tendon was transected. Ten rats were randomized to needling days 2-5. Mechanical testing on day 8 showed increased strength by 45% in the needling group. Next, another 24 rats were similarly unloaded, and 16 randomized to needling on day 5 after transection. Nineteen characteristic genes, known to be regulated by loading in this model, were analyzed by qRT-PCR. Four of these genes were regulated 15 min after needling. Three of them (Egr1, c-Fos, Rgs1) were among the five regulated genes after loading in a previous study. Sixteen of the 19 genes were regulated after 3 h, in the same way as after loading. In conclusion, needling increased strength, and there was a striking similarity between the gene expression response to needling and mechanical loading. This suggests that the response to loading in early tendon healing can, at least in part, be a response to microtrauma.

Place, publisher, year, edition, pages
American Physiological Society, 2014
Keywords
qRT-PCR; unloading; mechanical testing; early response genes; inflammation
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-103870 (URN)10.1152/japplphysiol.00741.2013 (DOI)000329196100008 ()
Available from: 2014-01-31 Created: 2014-01-30 Last updated: 2018-02-20
Eliasson, P., Andersson, T., Hammerman, M. & Aspenberg, P. (2013). Primary gene response to mechanical loading in healing rat Achilles tendons. Journal of applied physiology, 114(11), 1519-1526
Open this publication in new window or tab >>Primary gene response to mechanical loading in healing rat Achilles tendons
2013 (English)In: Journal of applied physiology, ISSN 8750-7587, E-ISSN 1522-1601, Vol. 114, no 11, p. 1519-1526Article in journal (Refereed) Published
Abstract [en]

Loading can stimulate tendon healing. In healing rat Achilles tendons, we have found more than 150 genes upregulated or downregulated 3 h after one loading episode. We hypothesized that these changes were preceded by a smaller number of regulatory genes and thus performed a microarray 15 min after a short loading episode, to capture the primary response to loading. We transected the Achilles tendon of 54 rats and allowed them to heal. The hind limbs were unloaded by tail-suspension during the entire experiment, except during the loading episode. The healing tendon tissue was analyzed by mechanical testing, microarray, and quantitative real-time polymerase chain reaction (qRT-PCR). Mechanical testing showed that 5 min of loading each day for 4 days created stronger tissue. The microarray analysis after one loading episode identified 15 regulated genes. Ten genes were analyzed in a repeat experiment with new rats using qRT-PCR. This confirmed the increased expression of four genes: early growth response 2 (Egr2), c-Fos, FosB, and regulation of G protein signaling 1 (Rgs1). The other genes were unaltered. We also analyzed the expression of early growth response 1 (Egr1), which is often coregulated with c-Fos or Egr2, and found that this was also increased after loading. Egr1, Egr2, c-Fos, and FosB are transcription factors that can be triggered by numerous stimuli. However, Egr1 and Egr2 are necessary for normal tendon development, and can induce ectopic expression of tendon markers. The five regulated genes appear to constitute a general activation machinery. The further development of gene regulation might depend on the tissue context.

Place, publisher, year, edition, pages
American Physiological Society, 2013
Keywords
tail-suspension; treadmill walking; tendon repair; early growth response; microarray
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-96124 (URN)10.1152/japplphysiol.01500.2012 (DOI)000319804600005 ()
Available from: 2013-08-14 Created: 2013-08-14 Last updated: 2017-12-06
Andersson, T., Eliasson, P., Hammerman, M., Sandberg, O. & Aspenberg, P. (2012). Low-level mechanical stimulation is sufficient to improve tendon healing in rats. Journal of applied physiology, 113(9), 1398-1402
Open this publication in new window or tab >>Low-level mechanical stimulation is sufficient to improve tendon healing in rats
Show others...
2012 (English)In: Journal of applied physiology, ISSN 8750-7587, E-ISSN 1522-1601, Vol. 113, no 9, p. 1398-1402Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
American Physiological Society, 2012
Keywords
Achilles tendon, mechanical stress, wound healing, hindlimb unloading, immobilization
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-85847 (URN)10.1152/japplphysiol.00491.2012 (DOI)000310649200007 ()
Note

Funding Agencies|Swedish Research Council|2009-6725|Linkoping University||Ostergotland County Council||King Gustaf V and Queen Victoria Free Mason Foundation||Swedish National Center for Research in Sports||

Available from: 2012-11-30 Created: 2012-11-30 Last updated: 2018-02-20
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