Currie JD, Kawaguchi A, Traspas RM, Schuez M, Chara O, Tanaka EM. Blastema formation and periosteal ossification in the regenerating adult mouse digit.

Can the authors correlate the spatial findings here on macrophage location with growth factor production (or lack thereof), as assessed in previous papers? Counts were calculated using the Cell Counter plugin for ImageJ (version 1.51d).
Similarly, msxb, which is a blastemal marker of epimorphic regeneration in the limb and fin, was expressed in the cells of the amputation plane (Figure 7). While we did not observe temporal differences in total macrophage accumulation between Acomys and Mus in the ear pinna, we did observe distinct differences in the spatial distribution of macrophage subtypes. Afterall, where’s the profit in self-healing? (I) Mallory’s triple-stained regenerate at 17 dpa showing the distal stump with newly differentiated woven bone (wb) that is capped with undifferentiated blastema cells (b). I was also thinking of creating a forum where people can connect and mingle with each other and sharing information. This cartilage disappeared as regeneration progressed (i, j). (D) Once the secondary amputation is complete, the wound epidermis closes through the region of degraded bone and a blastema forms. 2004 Jun 1;270(1):135-45. doi: 10.1016/j.ydbio.2004.02.016. These observations underscore the fluid nature of macrophage phenotypes in response to injury and disease and support a model where different subtypes differentially affect healing outcomes. In contrast, the cartilages that formed in the proximal amputation were maintained and fused with the Meckel's cartilages in the regenerate region. IBA-1 = red, DAPI = grey, autofluorescent red blood cells (RBC) = orange. Therefore, we investigated whether the regenerated cartilage formed in the distal amputation plane would be maintained when the plane remained in contact with the proximal amputation plane. Best of luck and do let me know what happens. I don’t mind being a guinea pig. For the BrdU‐labeling experiments, a 100 mM stock solution of BrdU (Wako, Osaka, Japan) dissolved in PBS was stored in the dark at −20°C.

Of the mammalian models of epimorphic regeneration that exist, very few have investigated how inflammatory cells affect blastema formation and regeneration. I am currently writing up a follow up article to this and Dr Cato T. Laurencin’s work is impressive.
The maximum value for each wound over the 15 images was used for subsequent analyses.

This fascinating subject requires the combination of the latest advances in regenerative medicine and tissue engineering. Total IBA-1+ area normalized to total DAPI+ area at D5 after the final treatment (J, n = 3 animals per treatment, *unpaired Student's t-test p<0.05). (M) At 28 dpa the regenerated woven bone (wb) has a mottled appearance and is distinct from the cortical bone of the stump. senescent cells). The dashed lines indicate the amputation plane.

In this light, their data supports our finding that M1 macrophages as marked with CD86 or F4/80 do not infiltrate the blastema. This observation is consistent with a previous report that periosteal cells differentiate into chondrocytes during mandibular bone regeneration in zebrafish (Paul et al., 2016). In response to this request we have amended our Discussion to include a possible role for senescent cells during regeneration based partly on Dr. Yun’s work in salamanders and newts, and on data from other regeneration models. After 1 week in culture, all BMDM were CD11b+ (Figure 5A). Images were collected on an Olympus BX51 upright microscope at 40x magnification.

Epimorphic regeneration proceeds with or without formation of a blastema, as observed for the limb and skin, respectively. The exception is if some of the nail bed is still intact.

Mus were fed mouse chow only. Our finding suggests that skeletal regeneration of the lower jaw mainly progresses through tissue regeneration that is dependent on the position in the jaw, and epimorphic regeneration plays an adjunctive role in this regeneration. (C) Histolysis of the stump is mediated by multinucleated osteoclasts (yellow) that erode the bone and creates a secondary amputation plane. We also isolated and activated BMDMs from Mus and confirmed they display a similar pattern of protein regulation in response to polarization (Figure 5J–R). In turn, blastema cells express several other genes; for example, muscle segment homeobox 1b (msxb) is a common marker of blastemal cells (Akimenko, Johnson, Westerfield, & Ekker, 1995; Nechiporuk & Keating, 2002). Similar gene expression was observed during lower jaw regeneration after amputation at the distal level (Figure S1). After sacrifice, femur and tibia were surgically removed, and mechanically cleared of all skin, muscle and tendon. Furthermore, the accumulation of senescent cells with age has been suggested to shorten lifespan, degrade tissue function, and increase the expression of pro-inflammatory cytokines in mammals (Baker et al., 2016, 2011). For luminol experiments: n = 7 Mus (14 ears) and n = 6 Acomys (12 ears) repeated measures ANOVA, F = 5.095, p<0.001 and for lucigenin experiments: n = 8 Mus (16 ears) and n = 6 Acomys, (12 ears), repeated measures ANOVA F = 4.536, p<0.001, *p<0.05 Sidak's multiple comparison test between species at the time indicated. A two-part list of links to download the article, or parts of the article, in various formats.


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