Prognostic Significance of Tumor-Associated Macrophages in Chondroblastoma and Their Association with Response to Adjuvant Radiotherapy

Figures & data

Table 1 Association Between TAMs Parameters and Clinicopathological Features of Chondroblastoma Patients

Factors	Categories	Number of Patients	CD68+ Cells Density		CD163+ Cells Density		CD163/CD68 Ratio
			Mean Level	P-value	Mean Level	P-value	Mean Level	P-value
Age (years)	Young (≤ cutoff^a)	105	527.2± 199.9	0.067	257.6 ± 100.8	0.429	0.52 ± 0.20	0.298
	Old (> cutoff^a)	27	606.7 ± 197.3		275.0 ± 103.7		0.48 ± 0.18
Gender	Female	46	544.0 ± 215.7	0.982	254.0 ± 96.4	0.550	0.50 ± 0.18	0.646
	Male	86	543.2 ± 194.3		265.0 ± 104.1		0.52 ± 0.20
Duration of symptoms (months)	Short (≤ cutoff^a)	118	536.8 ± 199.4	0.274	260.0 ± 99.6	0.710	0.52 ± 0.19	0.458
	Long (> cutoff^a)	14	599.3 ± 210.7		270.7 ± 117.4		0.49 ± 0.19
Tumor size (in diameter, cm)	Small (≤ cutoff^a)	28	486.1 ± 166.0	0.057	233.9 ± 84.2	0.073	0.51 ± 0.16	0.270
	Large (> cutoff^a)	104	558.9 ± 1207.7		268.5 ± 104.5		0.52 ± 0.20
Type of resection	EA	65	517.4 ± 206.5	0.144	254.3 ± 99.0	0.446	0.52 ± 0.17	0.635
	EI	67	568.7 ± 194.2		267.8 ± 103.6		0.51 ± 0.21
Surrounding tissue invasion	No	65	512.4 ± 187.0	0.081	231.7 ± 85.4	0.001	0.48 ± 0.16	0.053
	Yes	67	573.5 ± 211.1		289.7 ± 107.6		0.54 ± 0.22
Adjuvant radiotherapy	No	98	539.4 ± 200.3	0.746	262.2 ± 102.3	0.040	0.52 ± 0.20	0.021
	Ineffective	26	542.3 ± 208.1		282.3 ± 102.0		0.55 ± 0.17
	Effective	8	596.4 ± 208.5		179.3 ± 16.4		0.34 ± 0.12
Preoperative neurological dysfunction	No	97	544.2 ± 205.5	0.290	254.1 ± 100.6	0.181	0.49 ± 0.18	0.027
	Yes	35	513.6 ± 188.2		280.1 ± 101.9		0.58 ± 0.22
Postoperative neurological dysfunction	No	53	547.5 ± 197.9	0.851	242.0 ± 94.6	0.070	0.48 ± 0.19	0.081
	Yes	79	540.7 ± 204.6		274.0 ± 104.0		0.54 ± 0.19
Secondary ABC	No	74	548.4 ± 205.9	0.753	255.4 ± 95.9	0.472	0.50 ± 0.19	0.510
	Yes	58	537.2 ± 196.7		268.5 ± 108.0		0.53 ± 0.20
Chicken-wire calcification	No	63	532.5 ± 197.7	0.553	253.5 ± 100.7	0.407	0.50 ± 0.18	0.589
	Yes	69	533.4 ± 205.3		268.2 ± 102.0		0.52 ± 0.21
Tumor location	Axial	61	602.4 ± 212.8	0.002	322.5 ± 103.8	< 0.001	0.58 ± 0.23	< 0.001
	Extra-axial	71	492.8 ± 176.9		208.4 ± 61.9		0.46 ± 0.13
Tumoral p53 expression	Low (≤ cutoff^a)	101	544.9 ± 203.6	0.882	260.3 ± 102.3	0.861	0.51 ± 0.20	0.819
	High (> cutoff^a)	31	538.7 ± 196.4		264.0 ± 99.2		0.52 ± 0.18
Tumoral CD34 expression	Low (≤ cutoff^a)	77	551.4 ± 202.0	0.704	239.4 ± 90.5	0.004	0.48 ± 0.19	0.036
	High (> cutoff^a)	55	574.0 ± 205.9		291.7 ± 108.2		0.56 ± 0.19
Tumoral Ki-67 expression	Low	88	522.4 ± 195.6	0.090	242.8 ± 94.1	0.005	0.50 ± 0.18	0.156
	High	44	585.5 ± 208.0		297.9 ± 106.0		0.55 ± 0.22

Notes: Bold values indicate P < 0.05; ABC, aneurysmal bone cyst; p53, protein 53; TAMs, tumor-associated macrophages; ^aCutoff points were obtained by the “surv_cutpoint” function in the “survminer” package of R software. Cutoff points for patient age, duration of symptoms, tumor size, tumoral p53 expression, and tumoral CD34 expression in the survival analysis of OS were 38, 16, 2.0, 194, and 115 respectively.

Figure 1 Distribution of the tumor site for 132 chondroblastoma patients.

Figure 2 Representative images of tumor-associated macrophages parameters and immunohistochemical markers in chondroblastoma tissues.

Table 2 Univariate Analysis of the Prognostic Factors of Local Recurrence-Free Survival in Patients with Chondroblastoma

Factors	Categories	Number of Patients	Univariate Analysis
			χ^2	P-value
Age (years)	Young (≤ cutoff^a)	105	2.366	0.124^b
	Old (> cutoff^a)	27
Gender	Female	46	3.555	0.059
	Male	86
Duration of symptoms (months)	Short (≤ cutoff^a)	118	0.483	0.487^b
	Long (> cutoff^a)	14
Tumor size (in diameter, cm)	Small (≤ cutoff^a)	28	0.204	0.652^b
	Large (> cutoff^a)	104
Type of resection	EA	65	16.492	< 0.001
	EI	67
Surrounding tissue invasion	No	65	12.131	< 0.001
	Yes	67
Adjuvant radiotherapy	No	98	12.977	0.002
	Ineffective	26
	Effective	8
Preoperative neurological dysfunction	No	97	0.456	0.499
	Yes	35
Postoperative neurological dysfunction	No	53	2.860	0.091
	Yes	79
Secondary ABC	No	74	1.073	0.300
	Yes	58
Chicken-wire calcification	No	63	1.209	0.272
	Yes	69
Tumor location	Axial	61	3.388	0.066
	Extra-axial	71
Tumoral p53 expression	Low (≤ cutoff^a)	101	0.594	0.441^b
	High (> cutoff^a)	31
CD163+ cells density	Low (≤ cutoff^a)	103	24.834	< 0.001^b
	High (> cutoff^a)	29
CD68+ cells density	Low (≤ cutoff^a)	91	14.683	< 0.001^b
	High (> cutoff^a)	41
CD163/CD68 ratio	Low (≤ cutoff^a)	22	2.286	0.131^b
	High (> cutoff^a)	110
Tumoral CD34 expression	Low (≤ cutoff^a)	77	3.333	0.068^b
	High (> cutoff^a)	55
Tumoral Ki-67 expression	Low	88	1.661	0.197
	High	44

Notes: Bold values indicate P < 0.05; ABC, aneurysmal bone cyst; p53, protein 53; ^aCutoff points were obtained by the “surv_cutpoint” function in the “survminer” package of R software; Cutoff points for patient age, duration of symptoms, tumor size, tumoral p53 expression, tumoral CD34 expression, CD163+ cells density, CD68+ cells density, and CD163/CD68 ratio in the survival analysis of OS were 38, 16, 2.0, 194, 115, 392, 698, and 0.3, respectively. ^bP value from the Log rank test was corrected as previously suggested.

Figure 3 The relationship between tumor-associated macrophages parameters and response to radiotherapy in patients with chondroblastoma.

Figure 4 Kaplan–Meier curves of local recurrence-free survival of chondroblastoma patients stratified by CD68+ cell density and CD163+ cell density.

Figure 5 Multivariate Cox proportional hazard analyses of prognostic factors for local recurrence-free survival in patients with chondroblastoma. The results showed that type of surgery, densities of CD163+ and CD68+ cells were significant predictors of patients’ local recurrence-free survival (bold values used in the Figure indicate P < 0.05). Patients with Enneking inappropriate tumor resection and high number of CD163+ and CD68+ cells had a 4.124-, 2.929- and 2.236-times higher risk of chondroblastoma recurrence after surgery than their counterparts, respectively.

Table 3 Univariate Analysis of the Prognostic Factors of Overall Survival in Patients with Chondroblastoma

Factors	Categories	Number of Patients	Univariate Analysis
			χ^2	P-value
Age (years)	Young (≤ cutoff^a)	105	2.707	0.100^b
	Old (> cutoff^a)	27
Gender	Female	46	3.281	0.070
	Male	86
Duration of symptoms (months)	Short (≤ cutoff^a)	118	1.071	0.313^b
	Long (> cutoff^a)	14
Tumor size (in diameter, cm)	Small (≤ cutoff^a)	28	3.102	0.078^b
	Large (> cutoff^a)	104
Type of resection	EA	65	9.855	0.002
	EI	67
Surrounding tissue invasion	No	65	7.554	0.006
	Yes	67
Adjuvant radiotherapy	No	98	9.865	0.007
	Ineffective	26
	Effective	8
Preoperative neurological dysfunction	No	97	0.397	0.529
	Yes	35
Postoperative neurological dysfunction	No	53	8.002	0.005
	Yes	79
Secondary ABC	No	74	0.119	0.730
	Yes	58
Chicken-wire calcification	No	63	0.556	0.456
	Yes	69
Tumor location	Axial	61	0.018	0.893
	Extra-axial	71
Tumoral p53 expression	Low (≤ cutoff^a)	101	2.195	0.138^b
	High (> cutoff^a)	31
CD68+ cells density	Low (≤ cutoff^a)	91	15.310	< 0.001^b
	High (> cutoff^a)	41
CD163+ cells density	Low (≤ cutoff^a)	103	13.614	< 0.001^b
	High (> cutoff^a)	29
CD163/CD68 ratio	Low (≤ cutoff^a)	22	7.267	0.007^b
	High (> cutoff^a)	110
Tumoral CD34 expression	Low (≤ cutoff^a)	77	1.705	0.192^b
	High (> cutoff^a)	55
Tumoral Ki-67 expression	Low	88	0.582	0.445
	High	44

Notes: Bold values indicate P < 0.05; ABC, aneurysmal bone cyst; p53, protein 53; ^aCutoff points were obtained by the “surv_cutpoint” function in the “survminer” package of R software; Cutoff points for patient age, duration of symptoms, tumor size, tumoral p53 expression, tumoral CD34 expression, CD163+ cells density, CD68+ cells density, and CD163/CD68 ratio in the survival analysis of OS were 38, 16, 2.0, 194, 115, 392, 698, and 0.3, respectively. ^bP value from the Log rank test was corrected as previously suggested.

Figure 6 Kaplan–Meier curves of overall survival of chondroblastoma patients stratified by CD68+ cell density, CD163+ cell density and CD163/CD68 ratio.

Figure 7 Multivariate Cox proportional hazard analyses of prognostic factors for overall survival in patients with chondroblastoma. The results showed that type of surgery, density of CD163+ cells and CD163/CD68 ratio were significant predictors of patients’ overall survival (bold values used in the Figure indicate P < 0.05). Patients with Enneking inappropriate tumor resection and high number of CD163+ cells had a 3.286- and 6.354-times higher risk of death after surgery than their counterparts, respectively. However, patients with low CD163/CD68 ratio within the chondroblastoma microenvironment were 0.194 times less likely to have death following surgery than those harboring high CD163/CD68 ratio.