Effect of Calendula Officinalis Cream on Achilles Tendon Healing

In recent years, the scientific community has undertaken research on plant extracts, searching for compounds with pharmacological activities that can be used in diverse fields of medicine. Calendula officinalis L. is known to have antioxidant, anti‐inflammatory, antibacterial, and wound healing properties when used to treat skin burns. Therefore, the purpose of this study was to analyze the effects of C. officinalis on the initial phase of Achilles tendon healing. Wistar rats were separated in three groups: Calendula (Cal)—rats with a transected tendon were treated with topical applications of C. officinalis cream and then euthanized 7 days after injury; Control (C)—rats were treated with only vehicle after transection; and Normal (N)—rats without tenotomy. Higher concentrations of hydroxyproline (an indicator of total collagen) and non‐collagenous proteins were observed in the Cal group in relation to the C group. Zymography showed no difference in the amount of the isoforms of metalloproteinase‐2 and of metalloproteinase‐9, between C and Cal groups. Polarization microscopy images analysis showed that the Cal group presented a slightly higher birefringence compared with the C group. In sections of tendons stained with toluidine blue, the transected groups presented higher metachromasy as compared with the N group. Immunocytochemistry analysis for chondroitin‐6‐sulfate showed no difference between the C and Cal groups. In conclusion, the topical application of C. officinalis after tendon transection increases the concentrations of collagen and non‐collagenous proteins, as well as the collagen organization in the initial phase of healing. Anat Rec, 298:428–435, 2015. © 2014 Wiley Periodicals, Inc.

[1]  N. Maffulli,et al.  Ruptured Achilles tendons are significantly more degenerated than tendinopathic tendons. , 2001, Medicine and science in sports and exercise.

[2]  J. Carvalho,et al.  Arrabidaea chica extract improves gait recovery and changes collagen content during healing of the Achilles tendon. , 2013, Injury.

[3]  E. Hay,et al.  Guidebook to the extracellular matrix and adhesion proteins , 1995 .

[4]  L. Chiang,et al.  In vitro anti‐hepatoma activity of fifteen natural medicines from Canada , 2002, Phytotherapy research : PTR.

[5]  A. Oliveira,et al.  Pulsed LLLT improves tendon healing in rats: a biochemical, organizational, and functional evaluation , 2014, Lasers in Medical Science.

[6]  M. Simons,et al.  Auxiliary and autonomous proteoglycan signaling networks. , 2010, Methods in enzymology.

[7]  C. Puică,et al.  The hepatoprotective action of ten herbal extracts in CCl4 intoxicated liver , 2005, Phytotherapy research : PTR.

[8]  C. Rezende,et al.  Antifungal activity of the essential oil from Calendula officinalis L. (asteraceae) growing in Brazil , 2008, Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology].

[9]  D. Heinegård,et al.  [17] Isolation and characterization of proteoglycans , 1987 .

[10]  D. Buttle,et al.  Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. , 1986, Biochimica et biophysica acta.

[11]  C. Frank,et al.  Early medial collateral ligament scars have inferior creep behaviour , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[12]  S. Sosa,et al.  The role of triterpenoids in the topical anti-inflammatory activity of Calendula officinalis flowers. , 1994, Planta medica.

[13]  L. Soslowsky,et al.  Development of tendon structure and function: regulation of collagen fibrillogenesis. , 2005, Journal of musculoskeletal & neuronal interactions.

[14]  M. Mello,et al.  Optical anisotropy of collagen fibers of rat calcaneal tendons: An approach to spatially resolved supramolecular organization. , 2010, Acta histochemica.

[15]  D. Andreadis,et al.  Expression of matrix metalloproteinases 9 and 12 in actinic cheilitis. , 2013, World journal of experimental medicine.

[16]  G. Blandino,et al.  Antibacterial activity of medicinal plant extracts against periodontopathic bacteria , 2003, Phytotherapy research : PTR.

[17]  S L Woo,et al.  Injury and repair of ligaments and tendons. , 2000, Annual review of biomedical engineering.

[18]  R. Midura,et al.  Isolation and characterization of proteoglycans. , 1994, Methods in enzymology.

[19]  S. Khadabadi,et al.  Phytochemical constituents and pharmacological activities of Calendula officinalis Linn (Asteraceae): a review. , 2009 .

[20]  R. Kuttan,et al.  Effect of Calendula officinalis Flower Extract on Acute Phase Proteins, Antioxidant Defense Mechanism and Granuloma Formation During Thermal Burns , 2008, Journal of clinical biochemistry and nutrition.

[21]  O. Branford,et al.  The growth factors involved in flexor tendon repair and adhesion formation , 2014, The Journal of hand surgery, European volume.

[22]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[23]  Manuela Malatesta,et al.  Histological and Histochemical Methods - Theory and Practice , 2016, European journal of histochemistry : EJH.

[24]  G. Puddu,et al.  Overuse tendon injuries: Basic science and classification , 1997 .

[25]  Matthew J. Silva,et al.  The early inflammatory response after flexor tendon healing: A gene expression and histological analysis , 2014, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[26]  A. Oryan,et al.  A long term study on the role of exogenous human recombinant basic fibroblast growth factor on the superficial digital flexor tendon healing in rabbits. , 2011, Journal of musculoskeletal & neuronal interactions.

[27]  L. Galatz,et al.  The localized expression of extracellular matrix components in healing tendon insertion sites: an in situ hybridization study , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[28]  M. Mello,et al.  Proteoglycan arrangement in tendon collagen bundles. , 1984, Cellular and molecular biology.

[29]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[30]  N. Maffulli,et al.  Tenocytes from Ruptured and Tendinopathic Achilles Tendons Produce Greater Quantities of Type III Collagen than Tenocytes from Normal Achilles Tendons: An in Vitro Model of Human Tendon Healing , 2000, The American journal of sports medicine.

[31]  A. Okanlawon,et al.  A comparative study of the effects of bromelain and fresh pineapple juice on the early phase of healing in acute crush achilles tendon injury. , 2011, Journal of medicinal food.

[32]  N. Pavlova,et al.  Influence of the physiological regeneration and epithelialization using fractions isolated from Calendula officinalis. , 1982, Acta physiologica et pharmacologica Bulgarica.

[33]  L. Gulotta,et al.  Stem Cells for Augmenting Tendon Repair , 2011, Stem cells international.

[34]  B. de Campos Vidal,et al.  Experimental tendon repair: glycosaminoglycan arrangement in newly synthesized collagen fibers. , 2003, Cellular and molecular biology.

[35]  A. Goodship,et al.  The influence of ageing and exercise on tendon growth and degeneration--hypotheses for the initiation and prevention of strain-induced tendinopathies. , 2002, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[36]  B. Heinzmannb,et al.  etermination of the wound healing effect of Calendula extracts using the scratch ssay with 3 T 3 fibroblasts , 2009 .

[37]  R. Bank,et al.  Matrix metalloproteinase activities and their relationship with collagen remodelling in tendon pathology. , 2002, Matrix biology : journal of the International Society for Matrix Biology.

[38]  R. A. Rodrigues,et al.  Structural and biochemical alterations during the healing process of tendons treated with Aloe vera. , 2012, Life sciences.

[39]  N. Maffulli,et al.  Tendon healing: can it be optimised? , 2002, British journal of sports medicine.

[40]  Takashi Suzuki,et al.  Anti-inflammatory, anti-tumor-promoting, and cytotoxic activities of constituents of marigold (Calendula officinalis) flowers. , 2006, Journal of natural products.

[41]  G. Murrell,et al.  The Roles of Growth Factors in Tendon and Ligament Healing , 2003, Sports medicine.

[42]  A. Hensel,et al.  In vitro tests and ethnopharmacological investigations: wound healing as an example. , 2005, Journal of ethnopharmacology.

[43]  S. Hyslop,et al.  Organization of collagen bundles during tendon healing in rats treated with L-NAME , 2009, Cell and Tissue Research.

[44]  J. Trowbridge,et al.  Dermatan sulfate: new functions from an old glycosaminoglycan. , 2002, Glycobiology.

[45]  I. Merfort,et al.  Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblasts. , 2009, Journal of ethnopharmacology.

[46]  Nobuyuki Itoh,et al.  Novel 70-kDa Chondroitin Sulfate/Dermatan Sulfate Hybrid Chains with a Unique Heterogenous Sulfation Pattern from Shark Skin, Which Exhibit Neuritogenic Activity and Binding Activities for Growth Factors and Neurotrophic Factors* , 2005, Journal of Biological Chemistry.

[47]  H. Stegemann,et al.  Determination of hydroxyproline. , 1967, Clinica chimica acta; international journal of clinical chemistry.

[48]  D. Heinegård,et al.  Ultrastructural immunolocalization of cartilage oligomeric matrix protein (COMP) in relation to collagen fibrils in the equine tendon. , 2005, Matrix biology : journal of the International Society for Matrix Biology.

[49]  W. Lei,et al.  An experimental study on the effect of safflower yellow on tendon injury-repair in chickens. , 2011, The Journal of surgical research.

[50]  A. Tubaro,et al.  Topical Anti-Inflammatory Activity of Calendula officinalis Extracts , 1990 .